Draft Annual Report 2017

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INTERNATIONAL RUBBER

RESEARCH AND DEVELOPMENT

BOARD

Draft Annual Report for 2017

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Secretariat IRRDB, 260, Jalan Ampang, 50450 Kuala Lumpur, Malaysia International Rubber Research and Development Board (IRRDB) Overall Aims and Objectives: The IRRDB is a voluntary association of national organizations (Member Institutes) committed to research and development in natural rubber. The countries where these Institutes are located cover 96 per cent of world natural rubber production. The IRRDB conducts its business through a Board, responsible for all matters affecting the IRRDB; a Committee of Directors and Chief Executives - whose function is to examine, discuss and make recommendations on technical matters to the Board matters; and a Finance Committee. The IRRDB conducts a range of activities year-round including:

I. meetings to facilitate exchange of ideas between the staff of Member Institutes II. programmes initiated by the IRRDB’s Specialized Groups

III. training IV. International research programmes initiated by the IRRDB and executed by one or more

Member Institutes collaboratively, with occasional external funding. There are ten Specialized Groups, each operation being coordinated by a Liaison Officer: (i) Plant Breeding, (ii) Plant Protection, (iii) Physiology, (iv) Technology and End Uses, (v) Socio-Economic matters, (vi) Agronomy, (vii) Biotechnology, (viii) Exploitation Technology, (ix) Environment and (x) Transfer of Technology.

A copy of the IRRDB Constitution is available from the Secretariat (see below). Secretariat 260, Jalan Ampang, 50450 Kuala Lumpur, Malaysia Or P.O. Box 10150, 50908 Kuala Lumpur, Malaysia Secretary General Datuk Dr Abdul Aziz S.A. Kadir FASc Accountant Mrs. Nurul Syahidah Yunus Auditors Nasharuddin Wong & Co. Telephone +6(03)-42521612; +6(03)-92063750 Fax +6(03)-42560487 E-mail admin@theirrdb.org Website www.irrdb.com www.theirrdb.org

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THE BOARD

Chairman: Mr. Picheat Prommoon

Vice-Chairman: Vice-Chairman: Dr. Karyudi Prof. Zhou Jiannan

Cambodia Guatemala Myanmar

Mr. Ly Phalla Mr. Carlos Najera Mr. Hla Myint

Cameroon India Nigeria

Dr. Ehabe Eugene Ejolle Dr. A. Jayathilak Dr. A. I. Aigbodion

Mr. Jutheau D. Tousse Dr. James Jacob

China Philippines

Prof. Liu Guodao Indonesia Mr. Nicomedes Eleazar

Prof. Lin Weifu Dr. Chairil Anwar Mr. Rodolfo Galang

Prof. Zhou Jiannan Dr. Muhammad Supriadi

Dr. Karyudi Sri Lanka

Cote d'Ivoire Dr. Anang Gunawan Mr. N.V.T.A Weragoda Dr. W.M.G. Seneviratne Dr. Angiman Ackah Pierre

Dr. Yo Tiemoko Malaysia Thailand Mr. Picheat Prommoon

France Dr. Mohd Akbar Md. Said Dr. Napawan Lekawipat

Mr. Hubert Omont Mr. Tuan Mohamad Tuan Muda

Dr. Jerome Sainte-Beuve Dr. Zairossani Mohd Nor Vietnam Mr. Phan Thanh Dung

Mr. Do Kim Thanh

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Liaison Officers of IRRDB Specialist Groups

Plant Protection

Dr. Nguyen Anh Nghia Head, Crop Protection Division Rubber Research Institute of Vietnam HQ: 236bis Nam Ky Khoi Nghia, Dist.3, Ho Chi Minh City, Vietnam Lab: National Road 13, Lai Khe, Lai Hung, Bau Bang, Binh Duong, Vietnam Email: anhnghia@gmail.com; Mobile: +84 91 8340161

Plant Breeding

Dr.Kavitha K.Mydin, Joint Director (Crop Improvement) Rubber Research Institute of India Rubber Board P.O., Kottayam 686 009 Kerala, India. Email: kavitha@rubberboard.org.in

Socio-Economic

Wasana Wijesuriya PhD Principal Research OfficerA Biometry Section Rubber Research Institute of Sri Lanka Dartonfield Agalawatta 12200, Sri Lanka. Email: wijesuriyawasana@gmail.com

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Crop Management

Dr. Kuswanhadi Sembawa Research Center, Indonesia Rubber Research Institute. Jln Raya Palembang-Sekayu KM 29, P. O. Box 1127, Palembang 30001,Indonesia Tel : +62 (711) 361793 Email: kuswhd@yahoo.com

Molecular Biology & Physiology

Dr. Chow Keng See Principal Research Scientist (Functional Genomics) Production Development Division Malaysian Rubber Board Kuala Lumpur, Malaysia Tel: +603-6145 9598 Email: kschow@lgm.gov.my

Technology and End Uses

Mrs. Nopparat Vichitcholchai Rubber Technology Research & Development Division Rubber Authority of Thailand Ministry of Agriculture and Cooperatives 50 Phaholyothin Rd. Ladyao Chatuchak, Bangkok 10900, Thailand Email: processingsg@gmail.com

African Coordinator Dr. Eugene Ehabe Ejolle Inst. of Agricultural Research for Development (IRAD) P.O. Box : 2067 / 2123 Yaoundé Cameroon Email: eugene.ehabe@gmail.com

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Draft Annual Report 2017 – 2018

The Plant Protection Specialist Group

By

The IRRDB Liaison Officer for Plant Protection Specialist Group

Introduction

The Plant Protection Specialist Group is one of the eight (8) specialist groups of the IRRDB. The responsibility of the Plant Protection Specialist Group is to proactively monitor disease incidents in all NR producing countries, prepare quarterly status report for circulation to all member institutes and address other matters relating to rubber crop protection. The draft annual report 2017-2018 gathered and summarized the information on the progress of rubber plant protection activities from member institutes (Indonesia, Malaysia, Sri Lanka, Thailand, India, Cameroon, France, Myanmar, China and Vietnam).

Pest status survey of rubber plantations around the world has been ongoing. The progress of all activities of the IRRDB Plant Protection Special Group included:

ⅰ) Rubber protection activities of the IRRDB member countries and

ⅱ) Constructing the sharing and transfer platform of plant protection S & T.

Work tasks and progress in 2017

Pest status of rubber plantation in the world in 2017

More than 50 fungal pathogens and 20 insect pests have been identified as capable of attacking the rubber tree. Notably, the economically-important diseases and insect pests that they cause are:

the South American Leaf Blight (SALB), Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Corynespora Leaf Fall Disease (CLFD), Phytophthora leaf fall (PLF), Bark rot (BR), Root disease (RD), Bark beetles (Platypus secretus, P. calieutus, P. solidus, P. lewisi, Xyleborus aquilus, X. andrewsi, X. armipenis, X. fornicatus, X. noxius), Scale insect (Parasaissetia nigra) and Mites (Eotetranychus sexmaculatus, Tetranychus cinnabarinus, Eutetranychus orientalis, Oligonychus hainanensis, Brevipalpus huananis, Panonychus citri). Of these, only the SALB does not have a worldwide spread following efforts to control it.

However, in 2017, the economic importance of most traditional diseases and insect pests like Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Phytophthora leaf fall (PLF), Corynespora Leaf Fall Disease (CLFD), Root disease (RD), Bark beetles (Xyleborus aquilus), Scale insect (Parasaissetia nigra) and Mites (Eotetranychus sexmaculatus) were seen to have reduced in several rubber-growing countries following the production and planting of disease-tolerant clones.

Be that as it may, some of the new high-yielding breeds still succumb to new diseases and these have maladies spread to traditional clones.

Stem cankers caused by the fungus Lasiodiplodia theobromae in rubber plant is a new disease and has not yet been reported in Indonesia. (By Tri Rapani Febbiyanti)

A new leaf spot pathogen of Hevea brasiliensis was recorded in China. The causal pathogen was identified as Neofusicoccum sp., but conidia wa obviously less than other species of Neofusicoccum. The phylogeny of three molecular markers including ITS, EF-1a and β-Tubulin revealed that the causal pathogen was also significantly different from other species of Neofusicoccum. The causal pathogen was identified as a new species based on the morphological characteristics and molecular analysis. (By Huang Guixiu)

Leaf Blight in Cameroon caused by Pestalotiopsis microspora (By Aurelie I.C Nyaka Ngobisa)

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Table 1: Pest status of rubber plantation in several member countries in 2017

No Types Pests Status

1 Quarantine pests

South American Leaf Blight（SALB）

Restricted to tropical American countries and the rubber growing regions of Asia and Africa which remain as pest risk area (PRA)

2 Major Pests and Diseases

Corynespora Leaf Fall（CLFD） 2 to 5

White Root (Rigidoporus microporus) 3 to 11

Red Root（Ganoderma pseudoferreum Wakef） 4 or 12

Brown Root（Phellinus noxius (Corner) G.H.Cunn） 4 or 12

Purple Root（Helicobasidium compactum Boed.） 5 or 12

Stinking Root（Sphaerostille repens Berk. Et Br.） 5 or 12

Ustulina Root（Ustulina deusta (Hoffm.et Fr.)） 5 or 12

Black Root（Poria hypobrunnea Petch.） 5 or 12

Bark rot (BR) 4 to 12

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Recent origin or reached epidemic proportions of pests and diseases during the New Millennium

Fusicococcum Leaf Fall (Fusicoccum sp.) 5 or 12

Parasitic seed plant (Loranthus parasitica and

Macrosolen cochinchinensis（Lour）VanTeigh) 2 to 12

Gummosis (Botryodiplodia theobromae PAT.) 5 to 12

Patch canker/Basal rot (Fusarium solani) 5 to 12

Cockchafer grub attack 5 to 12

Patch canker/Basal rot 5 to 12

Sclerotium Collar rot of Hevea seedlings 5 to 12

Target leaf sport 5 to 12

Foot canker and sudden wilt 5 to 12

Papaya mealy bug 5 to 12

Bark beetles (Platypus sp., Xyleborus sp.) 4 to 12

Scale insect (Parasaissetia nigra) 4 to 12

mites ﹝Eotetranychus sexmaculatus Riley,

Tetranychus cinnabarinus（Boisduval）,

Eutetranychus orientalis（Klein）, Oligonychus

hainanensis Ma，Yuna et Lin）, Brevipalpus huananis

Ma et Yuna）, Panonychus citri（Mc Gregor)

4 to 12

4 Other Termite (Coptotermes curvignathus) 12

South American Leaf Blight（SALB） A five-day training workshop on South American Leaf Blight (SALB) of rubber for Asia and Pacific region was held in Brazil (13th – 17th November 2017). The training programme was led by NPPO Malaysia and co-organized by FAO and Michelin Brazil. It was proposed in the “Workshop To Develop Training programme and Reference Materials for Protection Against South American Leaf Blight (SALB) of Rubber”, which was held in Malacca, Malaysia in 2011. It was earlier scheduled in 2013 but was cancelled due to some unforeseen circumstances. However, as SALB is an important issue for the region and technical expertise on SALB is very critical, the APPPC Working Group on SALB proposed to have this hands-on training programme for personnel from the rubber growing countries within the Asia and Pacific region.

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Corynespora Leaf Fall Disease (CLFD) The most threatening rubber canopy disease in Asia and Africa caused by the fungus Corynespora cassiicola. This year, research projects have included education on epidemic surveillance, symptomatology identification, breeding for resistance and screening, development of effective spraying techniques as well as alternative disease management techniques (e.g. crown budding) which have been carried out in China, France, India, Malaysia, Indonesia and Sri Lanka.

Root diseases There are seven root diseases spread in rubber plantations since its domestication. Presently, white, red and brown root diseases have posed a threat in all rubber-growing countries in Africa and Asia except in India. The white root disease (WRRD) is the most threatening root disease in most rubber-growing countries, and red and brown root diseases are the most significant root diseases in China. In 2017, those several root diseases were controllable because of the biology of the pathogen. Its control has been further improved through early disease monitoring and detection systems, the development of resistant root stock and of downward translocating fungicides, the acceleration of stump decay as well as the use of effective biological control systems and antagonistic plant, etc.

Bark rot (BR) The main constraint in rubber plantations worldwide is the cessation of latex production caused by two different syndromes: (i) Tapping Panel Dryness (TPD) as a reversible physiological response to overexploitation; and (ii) Bark Necrosis (BN), an irreversible syndrome of the phloem that spreads from the collar. Bark Necrosis (BN) now affects most modern rubber plantations worldwide, with a wide range of severity across sites,depending on the clone, the age of the trees, the site and the country. Activities and proposed actions to control major pests and diseases in 2017 Strengthening the quarantine: South American Leaf Blight (SALB), Paracoccus marginatus. Monitoring and forecasting of plant disease and insect pests: Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Corynespora Leaf Fall Disease (CLFD), Eotetranychus sexmaculatus, Parasaissetia nigra and Xyleborus aquilus , etc. Biological and ecological characteristics of major pests and diseases. Developing pesticides for high - lift and its supporting techniques: Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Corynespora Leaf Fall Disease (CLFD) and Eotetranychus sexmaculatus , etc. Pesticide reduction technologies of rubber diseases and insect pests: Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Corynespora Leaf Fall Disease (CLFD), Bark rot (BR), Root diseases, Eotetranychus sexmaculatus, Parasaissetia nigra and Xyleborus aquilus , etc. Breeding for disease and insect pest resistance: Oidium leaf fall (OLF), Collectotrichum leaf diseases (CLD), Corynespora Leaf Fall Disease (CLFD) and Eotetranychus sexmaculatus , etc. General and specific training programmes for trainers and stakeholders (at international, regional and national levels).

Papers presented at the International rubber Conference 2017

1. Fetrina Oktavia (Indonesia). Analysis of Molecular Markers Linked to Corynespora Leaf Fall Disease Resistance in Rubber Plants.

2. Sekar Woelan (Indonesia). The Performance of IRR Series Rubber Clones on Trial Scale and in Commercial Plantations.

3. Syarifah Aini Pasaribu (Indonesia). The Resistance of IRR and RRII Series Rubber Clones to Leaf Fall Diseases in Indonesia and Its Correlation with Stomata Number & Cuticle Thickness.

4. Thakurdas Saha (India). Genetic Dissection of Disease Resistant Trait in Rubber through QTL Mapping

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5. Huang Guixiu (China) Opportunities and Challenges in Green Plant Protection Development of Natural Rubber in China.

6. Yu Zhang (China). Functional Analysis of HbMIol 2 in Powdery Mildew infection and Stress Response in Rubber Tree Hevea brasiliensis Muell Arg.

7. C Kuruvilla Jacob (India). Evaluation of Phylloplane, Cauloplane and Rhizosphere Bacteria for Multiple Antagonism Against Five Major Pathogens of Rubber (Hevea brasiliensis)

8. Nyaka Ngobisa (Cameroon). Characterization of Pestalotiopsis microspora, Causal Agent of Leaf Blight on Rubber (Hevea brasiliensis) in Cameroon.

9. Edison Purba (Indonesia). Distribution of Glyphosate-Resistance Goosegrass (Eleusine indica) Populations and an Alternative Herbicide (Indaziflam) to Control Them.

10. Hongli Luo (China). The Innate Immunity Responses in Rubber Tree. 11. Pham Thi Ngoc Giau (Vietnam). Biological Control of Corynespora cassiicola Causing Corynespora

Leaf Fall Disease (CLF) on Rubber Tree by Bacillus thuringiensis (T3) 12. Tri Rapani Febbiyanti (Indonesia). Pathogenicity Test of Lasiodiplodia theobromae Isolates from

Six Host Plants and Their Phylogeny Analysis 13. Alchemi P J Kusdiana (Indonesia). Resistance of Rubber Clones Recommended in Indonesia to

Corynespora and Colletotrichum Leaf Fall Diseases 14. Zaida Fairuzah (Indonesia). Bio-C : A New Biofungicide to Control Pink Disease (Corticium

salmonicolor) on Rubber Plants List of publications in 2017:

1. Ribeiro, S., Déon, M., Tran, D.M., Soumahoro, M., Masson, A., Clément-Demange, A., and Pujade Renaud, V. (2017). Functional analysis of cassiicolin, effector of the rubber tree pathogen Corynespora cassiicola. In 12th EFPP-10th SFP conference: Deepen knowledge in Plant Pathology for Innovative Agro-Ecology (Dunkerque, France).

Constructing the sharing and transfer platform of plant protection S & T in 2017

Five “one” Project construction has been going under the Belt and Road initiative.

A library: harmful organisms database of rubber tree.

A platform: pragmatic cooperation and exchanges on science, technology and biological resources and kept sound communication. Establish platform abroad, such as Experimental Station for Agriculture in Cambodian. With Sri Lanka to establish Tropical Agriculture R&D Park, to create a Sino-Cambodia Tropical Crops Demonstration Garden.

A book: The major problems involved in rubber trees protection in the world. A C\complete rubber plant protection of training textbooks in Chinese, English and French

A system: Effective transfer of technology system. Aid Myanmar and Cambodia to plant rubber. Affording technologies of breeding, high yield cultivation and so on, help Myanmar and Cambodia to develop rubber industry.

A network: Regional pest monitoring network.

New liaison officer and members of Plant Protection Specialist Groups At the meeting held on 22nd October, the IRRDB Board approved the recommendation of the Meeting of the Committee of Directors to appoint Dr. Nguyen Anh Nghia (Rubber Research Institute of Vietnam) as the Liaison Officer for the IRRDB Plant Protection Specialist Group from 23rd October 2017 to 22nd October 2020. Almost all member countries have nominated new representatives as listed in table 2:

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Table 2. List of members of Plant Protection Specialist Group (2017 -2020)

No Name and title Country No Name and title Country

1 Prof. Dr. Edson Luiz Furtado Brazil 10 Mr. Hla Myint Myanmar

2 Mr. Lim Khantiva Cambodia 11 Ms. Mi Khin Htay Than Myanmar

3 Prof. Fu Yueguan China 12 Dr. Victor Omorusi Nigeria

4 Dr. Valérie Pujade-Renaud France 13 Dr. Sarojani Fernando Sri Lanka

5 Dr. Shaji Philip India 14 Ms. Arom Rodesuchit Thailand

6 Dr.Tri Rapani Febbiyanti Indonesia 15 Dr. Nguyen Anh Nghia Vietnam

7 Dr. Wahounou Polie Jean Ivory Coast 16 Cameroon

8 Dr. Adam Malik Ahmad Zambri Malaysia 17 Guatemala

9 Mr Aizat Shamin Noran Malaysia 18 Philippines

Work tasks and progress in 2018

Pest status of rubber plantation in the world in 2018

A “pest survey on rubber tree” form has been sent to group members and responses from India, Indonesia, Thailand, Myanmar and Vietnam have been received. In general, SALB has not been detected in any Asian and African rubber-growing countries. CLFD is severe in India, Myanmar and Vietnam; mild in Indonesia and Thailand. OLF is severe in Myanmar and Vietnam; moderate in India, Indonesia and Thailand. CLD is severe in India; moderate in Indonesia, Myanmar and Vietnam; mild in Thailand. PLF is severe in India, Myanmar; moderate in Thailand and Vietnam; very mild in Indonesia. BR is severe in Myanmar and Vietnam; mild in Indonesia. Pink disease is severe in Myanmar and Vietnam; moderate in India; mild in Indonesia and Thailand. White root disease is severe in Thailand; moderate in Indonesia; mild in Myanmar. Some new diseases have been detected and have posed a more serious threat. These include Fusicoccum leaf blight in Indonesia. Previously, this disease was less important and considered an insignificant disease . Initially, the disease occurs in the area of North Sumatra since 2016, then spread to the Southern Sumatra region by the end of 2017 and spread until 2018. By March 2018, the disease developed into an epidemic, covering large areas of rubber plantations in that region with heavy defoliation. More than 25.000 hectares were attacked and spread in 7 provinces in Indonesia. Almost all the common clones were infected. This disease makes more than 60% to 90% of canopy defoliation with 15-25% reduction of latex. A leaf spot disease of Hevea in Malaysia caused by Pestalotia sp. has been detected in rubber plantations aged 10 -15 years in Johor State. It was found on clones RRIM 2001, RRIM 2025, RRIM 2023, PB 260, PB 350. The disease attacked leaves, shoots, twigs, fruits and seeds. The fungus has been isolated cultures and confirmation spores of Pestalotia sp. from infection of mature leaves of Hevea. In pathogenicity tests, tested isolates caused blight and necrosis on leaves. The pathogen was re-isolated from diseased leaves, confirming Koch's postulates.

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Activities and proposed actions to control major pests and diseases in 2018

Action on Fusicoccum Leaf Blight disease in Indonesia.

In April 2018, the IRRDB dispatched a team of experts comprising Dr. Kuruvilla Jacob, Dr. Adam Malik, Dr. Ramli Othman, Mr Tajuddin Ismail, Ms. Azimah Izhar, and Mr. Arif Makhdzir, a drone expert,to visit the affected areas to investigate and suggest control measures. Diseased leaf and root samples from severely affected trees were collected for further observation in the laboratory. The drone was used to evaluate the canopy density by collecting aerial images. The Plant Pathologists examined the samples collected from the fields. From these observations, it was clear that the causal organism of the leaf disease epidemic is Fusicoccum sp. (Neofusicoccum sp.). Based on the observations made, the experts confirmed that the epidemic is caused by Fusicoccum and suggested control strategies. A mission report was written by Dr. Kuruvilla Jacob. Besides, a report on present status of Fusicoccum leaf blight was also discussed and made by plant pathologists when they had a meeting in Palembang (Indonesia) in Aug. 2018.

International Plant Protection Workshop on Intergated Disease Management in Rubber Plantation.

This workshop was held at Aryaduta Hotel, Palembang, Indonesia from 31st July to 1st August 2018. The objective of the workshop is to update the research progress by the IRRDB research institute members on rubber disease management and also to discuss the Fusicoccum leaf disease outbreak in Indonesia recently. There were 147 participants from 14 countries (Cameroon, China, Côte d’Ivoire, India, Indonesia, Japan, Malaysia, Myanmar, Nigeria, Philippines, Sri Lanka, Singapore, Thailand and Vietnam). Eighteen (18) papers were presented in Keynote Address and three (3) sessions focused on: 1. Fusicoccum leaf disease; 2. Leaf disease; and 3. Stem and root disease.

Information exchange on Fusicoccum leaf disease and bark rot (Botryodiplodia theobromae)

Information on Fusicoccum leaf disease and bark rot (Botryodiplodia theobromae) was exchanged among group members.

In Indonesia, Fusicoccum disease become epidemic at the beginning of this year, in which bark rot was first detected.

In Malaysia Fusicoccum has long existed and bark rot has not been detected yet.

In Brazil, the Fusicoccum genus of fungi are known in Brazil to cause stem-end rot in various fruit plants, such as mango, guava, peach and avocado, but has not been detected in rubber tree.

In China, the leaf diseases caused by Fusicoccum were detected and found to have caused slight damage to rubber trees. The disease caused by Botryodiplodia theobromae is also present in China causing Black rotted bark with latex exuding and dieback of immature trees, with some cases of serious infestations in Hainan.

In Sri Lanka, Fusicoccum disease has not yet been detected. The bark rod is present in Sri Lanka. The invasion is mostly secondary. Any nursery or immature plant with the infections of Corynespora/Colletotrichum can get Botryodiplodia as a secondary invader. In mature plantations, Botryodiplodia causes bark cracks and sometimes cause knots as well. Here, if untreated during wet seasons, the result can be really destructive.

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In Vietnam, Fusicoccum disease has not been detected yet, while the disease caused by Botryodiplodia theobromae is common in almost all rubber planting areas. The Botryodiplodia disease could occur in nurseries, bud wood gardens, immature and mature rubber fielda.

The control measures were also been introduced and discussed.

Research activities Some specific researches are ongoing in IRRDB member institutes In Indonesia, studies to molecular identify the causative, epidemiology and control measure of Fusicoccum disease is implemented. In France, the characterization of Corynespora cassiicola effectors is in progress. Effectors are molecules secreted by one organism in order to manipulate the immune system of another organism and allow its colonization. It has been demonstrated that the deletion of the cassiicolin gene from the virulent Corynespora cassiicola isolate CCP causes a loss of virulence on the rubber tree. However, some moderately virulent isolates do not carry a cassiicolin gene, suggesting the existence of other effectors. Consequently, researchers developed a genomic/transcriptomic approach to identify in silico all putative effectors from CCP and other isolates representative of C. cassiicola genetic diversity. This molecular resource may provide interesting tools for effector-assisted selection of rubber trees with better tolerance to C. cassiicola. In Myanmar, three research projects are implementing: 1. Screening of Resistant Clones to Phytophthora Leaf Fall and Powdery Mildew (SLF) with the objective to assess the resistance of the PCRDC-bred clones to Phytophthora Leaf Fall and Powdery Mildew (SLF) and the use for clone selection and recommendation. On Phytophthora Leaf Fall (PLF), compared to BPM 24 (score 2), all the 8 tested PCRDC-bred clones are less resistant (score 3 - 4). Compared to PB 260 (score 3) which is moderately resistant to PLF, except one clone "PB 260 × RRIC 100(22)", the seven remaining tested PCRDC-bred clones are found to have the same resistance level (Score 3). "PB 260 × RRIC 100(22)" is found to be susceptible (score 4). On Powdery Mildew (SLF), disease severity varies according to the years, due to different micro-climatic situation during the refoliation period in those years. In the most severe year of 2014, compared to BPM 24 (score 4) which is severe and PB 260 (score 5) which is very severe, except "PB 260 × RRIC 100(22)", all the remaining 7 tested PCRDC-bred clones are found to have more or less similar severity levels. However, "PB 260 × RRIC 100(22)" is more resistant with (score 3). Therefore, "PB 260 × RRIC 100(22)" is moderately resistant to Powdery Mildew (SLF) and susceptible to Phytophthora Leaf Fall among the assessed clone; 2. “Preliminary Assessment of the Severity of Corynespora Leaf Fall Disease (CLFD) in Various Rubber Clones at PCRDC Nursery” with the objective to assess the severity of CLFD in various rubber clones planted in Myanmar. Among the 25 clones studied in PCRDC bud wood nursery, RRIC 110 is the only clone infected with CLFD having severity level (score 4). Between the two clones assessed in PCRDC polybag nursery, PB 260 is found to be more infected than BPM 24; and 3. “Preliminary Study on Antagonistic Effect of Trichoderma harzianum against the Pathogens of Some Rubber Leaf Diseases, in vitro” with the objective to determine the antagonistic effect of Trichoderma harzianum against the pathogen of rubber leaf diseases (Phytophthora, Corynespora and Colletotrichum) in vitro. It has been shown that Trichoderma harzianum could inhibit the growth of Phytophthora botryosa (58.30%), Corynespora cassiicola (59.09%) and Colletotrichum gloeosporioides (55.80%). Further studies and field trials need to be conducted on rubber nursery and immature planting. The anticipated finding would form a basis to formulate biological control measures to some extent on rubber leaf diseases.

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In Vietnam, a research project named “Study of Active Ingredients to Substitute Carbendazim in Control of Diseases on Rubber Tree (Hevea brasiliensis)” is being deployed with the aim of finding some active ingredients that can substitute carbendazim, which will be banned in Vietnam from 2019, as a move to control several popular diseases on rubber tree such as: Corynespora leaf fall (CLF), Oidium leaf fall (OLF), Colletotrichum leaf disease (CLD), Bird’s eye spot, Bark rot (BR) (Botryodiplodia theobromae). Some active ingredients belonging to triazole group and others are promising to substitute carbendazim in controlling diseases in rubber trees. Another project named “Set up website for Pest Diagnosis on Rubber Tree” is being set up in Vietnam at https://chandoanbenhonline.rubbergroup.vn/main.

Plant Protection Specialist Group Strategic Plan 2019 – 2023

The draft of the “Plant Protection Specialist Group Strategic Plan 2019 – 2023” was compiled and discussed at a meeting in Palembang held from 2nd – 3rd August 2018. The edited version was submitted to the IRRDB secretariat.

List of publications in 2018:

1. Lopez, D., Ribeiro, S., Label, P., Fumanal, B., Venisse, J.S., Kohler, A., de Oliveira, R.R., Labutti, K., Lipzen, A., Lail, K., Bauer, D., Ohm, R.A., Barry, K.W., Spatafora, J., Grigoriev, I.V., Martin, F.M., and Pujade-Renaud, V. (2018). Genome-Wide analysis of Corynespora cassiicola leaf fall disease putative effectors. Frontiers in microbiology 9:276. Papers to be presented at the IRRBB International Rubber Conference (will be supplementary)

1. Nguyen Anh Nghia, Vu Thị Quynh Chi, Nguyen Ngoc Mai, Nguyen Thi Kim Uyen, Nguyen Thi Thanh Trang, Đoan Nhan Luan, Vo Thi Thu Oanh, Nguyen Huu Khuong (2018) Study of Active Ingredients to Substitute Carbendazim in Control of Bird’s Eye Spot Disease (Drechslera heveae) on rubber seedling (Hevea brasiliensis).

2. Arom Rodesuchit, Somkid Dumnoi (2018) Efficacy and Methodology on Using N-fertilizers and sulfur Powder for White Root Disease Control in the Infectious Replanting Rubber Plantation.

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ANNUAL REPORT 2017-2018 Plant BREEDING SPECIALIST GROUP

THE IRRDB

Presented by Dr.Kavitha K.Mydin

Liaison Officer, Plant Breeding IRRDB

24 October 2018, Abidjan, Cote de Ivoire Contents

Sl. No Topic

A. Introduction : Major activities of the Plant Breeding Specialist Group in 2017-18

B. Report on Hevea breeding in IRRDB Member Institutes

1. Report on Hevea breeding activities in CRRI, Cambodia

2. Report on Hevea breeding activities in CATAS, China

3. Report on Hevea breeding activities in CNRA, Cote de Ivoire

4. Report on Hevea breeding activities in IRRI, Indonesia

5. Report on Hevea breeding activities in RRII, India

6. Report on Hevea breeding activities in Myanmar

7. Report on Hevea breeding activities in RRISL, Sri Lanka

8. Report on Hevea breeding activities in RRIT, Thailand

9. Report on Hevea breeding activities in RRIV, Vietnam

10. Annexure : Report on the IRRDB Plant Breeders’ Meeting 29-30 July 2018, Sembawa Research

Centre, IRRI, Indonesia

Annual Report 2017-2018 Plant Breeding Specialist Group PLANT BREEDING SPECIALIST GROUP The IRRDB Introduction : Major activities of the Plant Breeding Specialist Group Plant breeding activities in the year 2017-2018 in thirteen IRRDB member institutes were focussed on the Multilateral Clone Exchanges initiated in 2014. The countries that had imported clones were in the process of multiplication of the clones for raising source bushes or for laying out field trials. Those other countries still in the process of exchanging budwood are currently corresponding with one another on the matter. Some institutes that had already had the clones have initiated further studies towards the utilization of the clones for breeding. In addition to this very important multilateral exchange activity under the auspices of the IRRDB, the utilization of the 1981 wild germplasm for evolving improved clones was also underway. Each institute also had their own genetic improvement programmes on Hevea as per the priorities of the respective countries. The progress of research with respect to the IRRDB Multilateral Clone Exchange program, the 1981 IRRDB Wild Amazonian Germplasm and general Hevea breeding programmes of each institute are detailed in the forthcoming pages. The Plant Breeding Specialist Group met in the Sembawa Research Centre of the IRRI, Indonesia from 29-30 July 2018 to discuss details of the descriptors of the 49 clones exchanged under the 2016 IRRDB

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Multilateral Clone Exchange Program. A total of 22 Scientists from 10 countries participated in the deliberations and field visit. The purpose was to finalise the Compendium on the clones. A detailed report of the meeting is given in Annexure 1. The meeting provided an opportunity for the plant breeders to fine tune details of clone identification and the descriptors in the IRRDB format from eleven of the donor countries were completed. Since the clones contributed by CIRAD-Michelin have not been characterised as per the descriptors and since more original photographs would be required before printing the Compendium, it was decided to defer printing of the Compendium to a later date. Reports on the progress of the plant breeding programmes received from nine countries viz., Cambodia, China, Cote de Ivoire, Indonesia, India, Myanmar, Sri Lanka, Thailand and Vietnam under the IRRDB umbrella are presented below:

1. REPORT ON HEVEA BREEDING ACTIVITIES IN CRRI, CAMBODIA

Dr. Chetha Phen 1. The CRRII has 24 ongoing clone evaluation trials and has released clone recommendations

for 2018- 2020 as follows: 1. Recommendation for industrial rubber plantation:

Class I Class II Class III

IRCA 230 RRIM 600 PB 280 RRIC 121 IRCA 130

GT 1 IRCA 109 RRIM 712 PB 235 PB 217 RRIC 101 RRIC 100 KV 4

BPM 24 IRCA 111 IRCA 41 PR 303 PR 254 PB 310 PB 314 PB 324 PB 330 PR 261

2. Recommendation for small holders:

Clones Criteria

GT 1 Medium Productivity, Safe

RRIM 600 Medium Productivity, Safe

IRCA 230 High yield, Medium girth increment

RRIM 712 High yield, Medium girth increment

PB 280 High yield, good girth increment

Under the Multilateral Clone Exchange Program, the CRRI obtained a total of 13 clones from India, Thailand and Ghana. These have been multiplied and established in a budwood garden. Five of the CIRAD-Michelin clones were given in exchange to India and Thailand. During the year under report, no fresh exchanges were made, but correspondence on the matter is underway.

2. REPORT ON HEVEA BREEDING ACTIVITIES IN CATAS, CHINA

Mr. Zhang Xiaofei I. The Multilateral Clone Exchange program A total of 20 sets of budwood of 5 clones from CATAS were exported to the IRRDB member countries such as Thailand, India, the Philippines and Ghana. Additionally, 15 clones were

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introduced from Thailand, Ghana and India. These 15 clones have been placed for nursery clone trial and arranged for cold-resistant trial. II. The conservation and utilization of germplasm A total of 6,170 accessions of germplasm under 5 species and 1 variant of rubber tree were collected and conserved in the National Rubber Tree Germplasm Repository. There were 345 copies of excellent materials provided for research, teaching and production. III. Hevea breeding in China III.1. Crossing High-yielding clones Reyan106, Reyan88-13 and Reyan8-333, and cold-resistant materials IAN873, Tianren31-45 and the 1981’IRRDB germplasm were used as parents for crossing, 6 combinations were made via hand pollination, from which 550 seedlings were produced. III.2. Early identification of hand-pollinated generation Six seedlings were selected from 685 seedlings resulted from crossing in 2011, and their yield is more than 70g/t/t. Among the 102 clones under clonal trial 5 clones have good performance with an average yield of more than 20g/t/t. III.3. Large Scale Clone Trial The Clone Reyan 87-4-19 was established in 2006 and tapped in 2014. Its yield was 31.6 g/t/t at the first year of tapping, and 56.7 g/t/t at the fourth year of tapping. The RRIM 600 yielding 18.4g/t/t and 32.1g/t/t, respectively, was used as control. This clone had a girth growth of 7.6 cm before tapping and 3.6 cm per year after tapping, while the RRIM 600 as the control had a girth growth of 6.7 cm and 3.4 cm, respectively.

III.4. New variety demonstration More than 200 ha of rubber plantations of clones such as Reyan7-20-59, 8-79, and 7-33-97 were established for demonstration in different ecozones in China during the periods of between 2010 and 2017. The Reyan 8-79 had a good performance at Mengding Farm in Yunnan province, and yielded 2.4 kg/t/t, 4.3 kg/t/t, 5.9 kg/t/t, 7.1 kg/t/t, 7.6 kg/t/t, 9.5 kg/t/t, and 11.2 kg/t/t at the year of tapping from the first to the seventh year.

3. REPORT ON HEVEA BREEDING ACTIVITIES IN CNRA, COTE DE IVOIRE Dr. Angeline Elabo The IRRDB clone exchange by multilateral agreement between Côte d’Ivoire and other countries involved in the project began in March 2016 and is currently in progress. To date, Côte d'Ivoire through the CNRA has received and met demands of IRCA clones from the Philippines, India, Thailand and Sri Lanka. It is expecting to receive import permits from other countries so as to process and send to them IRCA clone budwoods. Over the 44 expected, 16 clones from the Philippines (USM1), India (RRII430, RRII417, RRII414, RRII422, RRII429), Thailand (RRIT408, RRIT251, RRIT226, RRIT3604, RRIT39043) and CIRAD Ghana (FDR5788, FDR5665, FDR4575, CDC312) were introduced in Côte d'Ivoire.

4. REPORT ON HEVEA BREEDING ACTIVITIES IN IRRI, INDONESIA Dr. Fetrina Oktavia

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I. Status of multilateral clone exchange in Indonesia Multilateral Clone Exchange of IRRDB Members

No. Institution Import Export Remark

1. Rubber Research Institute of India (RRII)

Done 2015 (5 clones)

Done 2015 (3 clones)

Multiplied in Sungei Putih Research Centre - IRRI

2. Philippine Rubber Research Institute (PRRI)

Process in 2018 (1 clone)

Done 2018 (3 clones)

Multiplied in Sungei Putih Research Centre - IRRI

3. Rubber Research Institute of Thailand (RRIT – RAOT)

Done, but can’t get it, problem in a customs,2018 (5 clones)

Done 2018 (3 clones)

Multiplied in Sembawa Research Centre - IRRI

4. Ghana Rubber Estate (CIRAD)

Process in 2018 (5 clones)

Process in 2018 (3 clones)

Multiplied in Sembawa Research Centre - IRRI

5. Centre National de Recherce Agronomique (CNRA)

Not Respond - -

Pedigree of Indonesian clones given in exchange under MCE

II. Status of maintenance and utilization of 1981 germplasm The maintenance of IRRDB 1981 germplasm is in the budwood garden. The germplasm are maintained at Sungei Putih and Sembawa Research Centre, IRRI. Some of the genotypes were used as parents on the breeding program. There are several studies related to IRRDB germplasm:

1. Resistance Identification of IRRDB 1981 Rubber Germplasm to Corynespora Leaf Fall Desease Based on Cassiicolline Toxin Activity

One of the most important diseases of rubber tree is Corynespora Leaf Fall (CFL) disease, caused by the fungus Corynespora cassiicola. The pathogen can attack through all of rubber plant growth stages, leading to a significant decrease of latex production and even causing plant death. Using resistant clones as a planting material is the most effective and economical way to prevent CLF disease. Therefore, identification of resistant clones has become the main strategy in the management of CLF disease. This study aims to identify the degree of resistance of the 50 selected accessions of IRRDB 1981 (PN'81) germplasm to CLF. The 6 clones of Wickham population were used as a control (BPM 24, BPM 1, GT 1, RRIC 600, PB 260 and RRIM 600). Four isolates of C. cassiicola (CC-01, CC-20, CC-22, and CC-23) were inoculated to young leaves of B2C stage of

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growth independently, and the intensity of wilting leaves was calculated based on water losses estimation due to the pathogenic-derived toxin activity. The results showed that 12 accessions were very resistant, 13 accessions were resistant, 23 accessions were susceptible and 8 accessions were very susceptible. The PN 451, PN 494 and PN 604 accessions showed better survival rates against CLF compared Wickham population which these accessions might potential to be used as a resistant gene source in the rubber breeding program.

2. Genetic Diversity and Population Structure of IRDB 1981 Germplasm Based on EST-SSR

The objective of this study was to reveal the genetic diversity and population structure of rubber germplasm consisting of 50 accessions of IRRDB 1981 germplasm and 6 clones of Wickham populations using 15 pairs of EST-SSR primers. The result showed that all primers used in this study generated polymorphic markers and the SSRH-268 primer pairs showed the highest value of population N, PIC, Ho and He, respectively. Phylogenetic tree based on matrix dissimilarity simple matching and constructed using Neighbour Joining method showed that the evaluated materials belonged to three clusters with less admixture genotype on each clusters. The Wickham population belonged to the different group than that of IRRDB 1981 ones. Such results indicate that Wickam and IRRDB rubber accessions have different genetic background. The IRRDB 1981 accessions were further divided into two sub-clusters, in which the first cluster was dominated by Rondonia accessions and the second cluster was dominated by Mato Grosso accessions and adimixture partly of accessions from Rondonia. Althought it has not completely divide the germplasm into different cluster, but the clustering mostly in lines with the geographical origin of the evaluated materials. Distribute population based on PcoA further support the results of cluster analysis and indicated that Rondonia and Mato Grosso more divergence compare others, Acre closely to Rondonia and Wickham separated from IRRDB 1981. The population structure analysis mostly in line with cluster analysis and PcoA demonstrated the presence of three population structures of the rubber germplasm. Results of the genetic diversity and population structure analysis would be useful for future rubber breeding analysis, such as in selecting clones for parents for hybridization programmes and for conserving genetic resources of rubber germplasm. III. General Hevea breeding activities in IRRI 1. Hybridization and Genotype Selection

The study aims to obtain new superior genotypes that are more productive with dry rubber yield potential > 3 t / ha/ year and has an important and better secondary characteristic, which is resistant to leaf fall disease and form a light canopy. Cultivars with a prototype rubber with the productivity per unit area, is maintained during the economic cycle of the rubber plant. The hybridization was conducted at the pollination garden of Sungei Putih and Sembawa Research Centre, IRRI. Research methods were carried out by crossing between of Hevea parents which are genetically distant, in order to obtain large diversity and heterosis in their derivatives. The combination of parent crossing are clones of PB 217, PB 260, PB 330, PB 340, IRR 39, IRR 42, IRR 112, IRR 118, IRR 220 as a female parent and clones of PB 260, PB 330, PB 340, RRIC 131, IRR 42, IRR 111, IRR 219, IRR 220, PN 462, PN 389, ORTET 7106, ORTET 7684 as a male parent. Observations conducted on the percentage of fruitset, the number of seeds produced, and the number of plants (F1). The number of crosses on 2017 was as much as 22429 flowers, with 5% of fruitsets, 2.3% of seeds and 0.3 of F1 plants. The evaluation of the F1 population on SET hasbeen done on girth, barkthickness, plant height, first branch height and the potential production of dry rubber.

2. Trial of Clones Rubber Promising of IRR Series 300 and 400 on Different Enviromental

Condition The election of IRR series clones is carried out systematically from the Promotion Plot Trial, Small Scale Clone Trial, Large Scale Clone Trial and Adaptation Trial. The clones selected are

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then recommended as new superior and promising. The evaluation of the performance of promising clones at commercial planting is to be carried out by integrating with other disciplines. Some of the IRR 300 and 400 series clones were evaluated on different of climate conditions. Some clones showed vigorous growth and high production on SSCT, Promotion Plot Trials and LSCT. The selection of IRR 300 series clones that consist of IRR 307, IRR309 and IRR 319 clone have a girth of about 45 cm in 4 years and average rubber yield was > 48 g/t/t when tapped for 7 years. These clones are resistant to Colletotrichum and Corynespora and moderately resistant to Oidium leaf fall diseases. Clones IRR 425 and IRR 429 have a girth of about 49.0 cm when 4-year old. The clones have good yield potential with an average rubber yield about > 55 g/t/t on 5 years of tapping. The development of these clones on the commercial scale could be done gradually by considering their suitability for different plant growth environments.

3. Genome and Genetic Analysis of Tapping Panel Dryness on Rubber The loss of rubber production of up to 20% caused by Tapping Panel Dryness (TPD) is one of

the most serious threats on rubber cultivation. The occurrence of TPD which had been mostly found on the quick starter clones and conventional breeding programme, often led to the selection of the quick starter clones with high susceptibility to TPD and others abiotic stress. Due to this problem, the Indonesian Rubber Research Institute (IRRI) and CIRAD, France have set up a collaborative research activity with the aims of carrying out genetic analysis of the segregating population for mapping and QTL analysis linked to TPD susceptibility, the identification of genes correlated to TPD underlying of QTLs, comprehensive analysis of latex production and TPD, analysis of genetic variability correlated to water stress, and the selection of the best genotypes. The trial was established in November 2016 where the planting at standard density (6 x 3 m) on 18 ha area was carried out. The planting material consists of 202 legitimated segregating population genotypes from the parents PB 260 (susceptibility to TPD) and SP 217 (tolerant to TPD), the parents clones and 6 control commercial clones (GT 1, AVROS 2037, IRR 112, IRR 39, PR 261, RRIC 100). All the population had been tested for legibility by SSR markers to ensure that the planting progeny is the true result of crossing PB 260 and SP 217 and to avoid any mistakes on the multiplication process.

These progenies were obtained from the crossing of parent on 2012 to 2014 at the pollination garden of Sembawa Reserach Centre, IRRI. The progenies were planted on SET as a source of budwood for multiplication. The growth data collection and analysis of resistance to disease (white root disease, Corynespora, Colletotrichum, Oidium and Fusicoccum leaf fall diseases) were done during the immature period. Additionally, data of climate factors such as soil moisture, rainfall and number of rainy days was recorded. The observation of one-year old plant showed that these genotypes growth variation with a girth of about 7.3 – 15. 3 cm. Observation and laboratory analysis showed that these genotypes are resistant to Corynespora and Colletotrichum leaf fall disease.

5. REPORT ON HEVEA BREEDING ACTIVITIES IN RRII, INDIA Dr.Kavitha K. Mydin Bridging the escalating gap between demand and production of natural rubber is expected to drive greater expansion of plantations in the non-traditional regions of India. Since this would require the movement of clones to newer environs, there was an urgent need for the identification of locally adaptive region-specific genotypes. Envisioning of this vital necessity led to a significant breakthrough on region-specific clone recommendation. Exotic clones of Brazilian origin, harbouring potential disease resistant genes, were imported and added to the Hevea gene pool of the country, providing ample scope for their utilization in breeding for productive as well as adaptive traits. Clones imported under the international clone exchange

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of 2014-17 were screened for tolerance to biotic and abiotic stress parameters. Conventional genetic improvement including hybridization, polycross breeding and ortet selection continued with the aim of developing clones with high rubber yield and important secondary traits. Ninety superior pipeline clones were under evaluation in Central Large Scale Trials and satellite on-farm trials spread over 44 locations in traditional and non-traditional regions for selecting superior clones through the participatory plant breeding approach. Pipeline clones were also evaluated for their response to Ethephon stimulation.

I. International Clone Exchange under the IRRDB MCE Program. The replenishment of domesticated germplasm with clones from other rubber growing countries via bilateral and multilateral exchanges was successfully achieved. A total of 44 high yielding and disease resistant clones have been imported thus from 10 countries from 2016 onwards. During 2017, five CMS SALB -resistant clones viz. FDR 5597, FDR 5802, CDC 56, MDX 624 and CD 1174 were imported from Ghana and two CMS SALB-resistant clones viz., FDR 4575 and FDR 5665 were imported from Cambodia. The above exotic clones were successfully recovered through budding and multiplied and were subsequently planted in source bush nurseries at the Central Experimental Station, Chethackal. Four clones viz., IRCA 41, IRCA 733, IRCA 347 and IRCA 825 were re-imported in 2017 from Côte d'Ivoire. While there was very low budding success for three of the clones, one clone did not show sprouting. Disease screening in the Source Bush Nursery at the Central Experiment Station of RRII showed that no major disease incidences were observed in IRCA 331, two ARCPC clones from Myanmar and clones imported from other countries including Vietnam, China and Cambodia. Three clones (USM1, Philippines; RRISL 203 and RRISL 219, Sri Lanka) were also imported. The above clones were bud-grafted and a polybag nursery was established at CES. Clones imported from other countries including those from Vietnam (8), China (5), Cambodia (5), Thailand (5), Indonesia (3) and Sri Lankan (3) were also maintained in the SBNs at CES. The imported clones in the International Hevea Clone Museum (2017) were screened for tolerance to Corynespora through toxin-bioassay in collaboration with Pathologists. The clones Baoting 936 and RRIT 226 showed relative tolerance against Corynespora during in vitro leaf wilt bio assay using crude toxin In order to assess the drought tolerant potential of some of the recently introduced exotic clones, a few crucial physiological parameters were tested in vitro as well as in the field grown plants. The screening programincluded the study of parameters like photosynthetic CO2

assimilation rate , transpiration rate, stomatal conductance , chlorophyll fluorescence and cell membrane stability. Among the various parameters tested, there was no single clone that outperformed all others in terms of stress tolerance. Based on the results from different experiments, the clones are graded for degree of drought tolerance as follows: Better tolerant clones: RRIT 226, RRIV 106, RRIT 251, RRIV 5, RRIV 1, RRIT 3604 Moderately tolerant: RRIT 3904, FDR 4575, RRIT 408, RRIV 103, RRIV 114, RRIV 124 Poorly tolerant / relatively susceptible: Baoting 936, Reyan 7-20-59, CATAS 7-33-97, RRIV 109, RRIV 106, FDR 5665 A study on genetic divergence among the MCE clones using molecular markers was initiated in 2017. Out of the 44 clones imported, 34 clones along with 10 other popular clones including

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two clones among the RRII 400 series were included in the study. DNA fingerprinting of the clones from Indonesia, France, China, Thailand, Philippines, Vietnam, Sri Lanka, India, Myanmar and Cote d’ Ivoire was carried out using simple sequence repeat (SSR) markers to investigate genetic variations and relationships among these clones. Twenty-three SSR markers generated 107 alleles in the study. The clones were found to be highly diverged as genetic distance ranged from 30% to 70%, forming 15 clusters at a distance coefficient of 0.5. Cluster analysis clearly separated clones of Indian origin. Among the 34 clones 7 clones belonging to CIRAD-Michelin and received from Cambodia (4 clones) and Ghana (2 clones) showed diversity >60%. Clone FDR 5802 representing a separate cluster showed the maximum diversity of >70%. Out of 34 clones, 12 clones could be identified based on unique profiles generated by these SSR markers. Thus, clone specific SSR profiles generated could be used for their identity. An international Hevea clone museum, consisting of high-yielding as well as SALB-resistant clones imported from various countries was established and maintained at RRII Main Campus, Kottayam.

II. Conservation and utilization of the 1981 IRRDB Wild Germplasm in India

The narrow genetic base of cultivated rubber in the east which was causing yield improvement levels in breeding programmes to plateau, as well as the fact that the primary center of diversity and richest source of variability – the Amazon forests, were disappearing rapidly made it highly imperative to collect and conserve all available germplasm at the earliest. The highly successful wild germplasm collection expedition in 1981 by the IRRDB in collaboration with the EMBRAPA into Acre, Rondonia and Mato Grosso in Brazil resulted in a huge collection that was eventually shared by the member countries. India’s share of the wild germplasm was introduced into the country from the Malaysian-based nursery between 1984 and 1990 in batches, and is being conserved at two locations – one in the traditional region in Kerala, and the other in the non-traditional region of the northeast. Conservation A total of 4548 accessions was finally established in the nurseries, 3576 and 972 accessions in Kerala, South India and in three states in North East India respectively. This ex situ conservation is mainly in the form of source bushes. Arboretum: A programme to conserve the entire Hevea germplasm available in India, both wild and Wickham, in the form of an arboretum at Teksragre in Tura, Meghalaya, in the north-east of India was initiated in 2014, with the intention of establishing an insurance collection. More importantly, as the collection will comprise the 1981 IRRDB wild germplasm interspersed with the Wickham clones, panmixis in this population will also result in natural genetic mixing between the accessions from four different locations – Acre, Rondonia and Mato Grosso, as well as Manaus from where the original Wickham collection had been made. This will ensure a regular and easy supply of open pollinated seeds with new recombinants every year for evaluation, without the need for manual hybridisation. Simultaneously however, hybridisation between selected accessions will also be possible whenever necessary. So far, a total of 466 wild Amazonian and Wickham accessions have been established here. Another arboretum established earlier and comprising of 120 accessions, is being maintained at Central Experiment Station (CES), Chethackal of RRII.

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Evaluation Characterization and evaluation of the wild germplasm are essential prerequisites for its utilisation. However, in view of the size of the collection and the perennial nature of the crop, this can be done only in phases as per availability of land and manpower resources. Preliminary Evaluation Nine relatively high yielding accessions AC 3131, AC 552, RO 2136, RO 1313, AC 567, AC 1964, RO 341, MT 4351 and RO 210, and vigorous accessions for timber traits MT 4219, AC 4140, MT 387, AC 647 and RO 2883 have been identified from the preliminary evaluation in nurseries. A comprehensive preliminary evaluation in the juvenile phase of the entire collection in a short time became possible in the newly re-established SBNs as they provide uniform sets of accessions in terms of age. As the accessions have now been planted at a spacing of 1 x 1 m, in an augmented RBD, with check clones and a plot size of five, morphological characterization and preliminary evaluation in the juvenile stage for the entire collection was possible here. However, since these nurseries have been planted at close spacing and had to be cut back after 4 years after juvenile stage data collection, only those selected accessions that go on to the full-fledged evaluation trials can be characterized in the mature phase. The entire collection here has been morphologically characterized for qualitative traits in the juvenile stage according to the descriptor prepared by Rao et al. (2005). The entire set has also been subjected to preliminary agronomic evaluation, including test tapping in the 3rd and 4th years of growth before being cut back for budwood. Field tolerance to drought was also assessed here by visual scoring for yellowing and senescence. Preliminary selections identified for vigour, test tap yield, latex vessel number etc. are being put into detailed further evaluation trials (FETs)

As expected, preliminary evaluation through test tapping of the entire collection showed that most of the wild accessions are low yielders compared to the popular Wickham clones. However, a few individual accessions with comparable or even higher yield than the popular clone RRII 105 have been identified. Wide variability and significant differences among the accessions for most of the agro-morphologic traits, bark structural characters and juvenile yield indicated the scope for selection of accessions with desirable characters. All these accessions are to be evaluated in detail in statistically laid out further evaluation trials to confirm their yield potential. Profiling of bark anatomical traits has identified six wild accessions (MT 1033(7.9), RO 399(7.8), OR 1177 (8.1), AC 2646 (8.6), RO 3000 (7.2) and RO 2946 (7.7) with a high number of latex vessel rows and these have been included in the breeding pool for yield improvement.

At present, apart from the preliminary selections which are already under further evaluation, 81 wild accessions with more than 80% of the test tap yield of the control and another 61 accessions with 50-80% of the control’s test tap yield, identified from various preliminary studies so far, are to be evaluated further. Further evaluation and selection

Accessions that perform well in preliminary evaluations are taken for further evaluation in full-fledged field trials: those accessions showing 80 per cent or more test tap yield than the check clones are planted in statistically laid out further evaluation trials (FETs), while those with 50-80 per cent of the yield of the check clones are evaluated first in clonal nurseries (CNs). These accessions are also being used in hybridization programmes to infuse new alleles into the gene pool.

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a. Clonal nursery evaluation

A clonal nursery comprising 15 selected wild accessions from SBN 2004 having 50- 80% test tap yield of RRII 105 was planted at CES, Chethackal along with three check clones at a spacing of 2.5 x 2.5 m for evaluating their yield potential. After two rounds of test tapping, three accessions (AC 2199, MT 1056, and AC 2027) with good growth and test tap yield higher than RRII 105 were identified. They were selected for future crop improvement programmes.

b. Further Evaluation Trials There are currently six FETs comprising 117 accessions planted at three different locations. The first FET in 1995 comprising 80 wild accessions preliminarily selected on the basis of test tap yield and timber potential from nursery studies, was completed. AC 166 identified from this trial has been forwarded to on-farm testing in 5 locations in Kerala. The other selections from this trial include AC 2004, RO 2385 and RO 2908 with 85 -102 % the yield of the high yielding check Wickham clone RRII 105. Potential high timber yielding accessions are listed in Table 1: Table 1. Potential accessions for timber yield (16th Year)

Accessions

Timber yield (bole volume m3)

Dry rubber yield (g/t/t)

MT 941 0.16 15.38 (24.0%)

RO 1347 0.14 14.78 (23.0%)

MT 199 0.14 8.94 (13.9%)

MT 1630 0.14 5.33 (8.3%)

AC 661 0.14 2.01 (3.13%)

RO 255 0.13 24.70 (38.6%)

MT 915 0.13 18.88 (29.5%)

MT 68 0.13 2.95 (4.6%)

MT 1032 0.13 10.97 ( 17.1%)

AC 3013 0.13 32.74 ( 51.1%)

RRII 105 0.08 64.04

CD(P= 0.05) 0.04 20.06

* Figures in parenthesis : % of the yield of the check clone, RRII 105 In FET 2003, 22 preliminary selections of wild Hevea accessions along with three modern clones are being evaluated in the traditional rubber growing region of India. In another FET planted in 2005 with 22 preliminary selections, 2 accessions (AC 2004 and MT 4788) show yield on par with the control in the second year of tapping, while another had 80% of the yield of the control. At Padiyoor in North Kerala, 19 wild accessions were evaluated for latex-timber traits. At the age of 11 years, timber volume was significantly higher in 2 wild accessions (MT 941 and AC 650). Two accessions (AC 685 and AC 707) recorded yield of 28.3 and 30.3 g/t/t, which were statistically on par with RRII 105 and PB 235 (38.7 and 30.5 g/t/t ). Three more sets of 26, 13 and 22 selections from the preliminary evaluation in the reestablished SBNs have been planted in FETs in 2008, 2010 and 2013 respectively at CES, and another 12 at RRS, Dapchari in the drought prone tract in 2014.

On-Farm evaluation:

The accession AC 166 has been forwarded to on-farm evaluation (OFT) in 5 estates in 2010, considering its consistent good yield performance in Further Evaluation Trial. The performance of this accession in terms of growth after eight years has been good compared to other test clones so far.

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Screening for stress tolerance a. Biotic stress resistance (Disease) For Phytophthora, out of 152 accessions short listed, 18 were found to have tolerance level equal or better than RRII 105. Among these 18, 14 accessions represented Mato Grosso provenance indicating genetic potential of MT accessions towards Phytophthora tolerance, as reported earlier also. In the further evaluation of 80 wild accessions in the mature stage, it was observed that accession MT 922 had the highest leaf retention (82.86 %) followed by MT 999 (76.67%) whereas it was only 60% in the susceptible clone RRII 105. This shows the tolerance potential of these two accessions against Phytophthora infestation. A set of 41 shorted listed wild Hevea accessions along with 2 control clones (RRII 105 and GT 1) are under evaluation for confirmation of field tolerance to Corynespora in a hotspot area for the disease. b. Abiotic stress resistance (Drought & Cold) Field screening of the wild accessions in the drought prone areas in Dapchari (Maharashtra), and in the cold sub-Himalayan region at Nagrakatta (Northern part of W. Bengal), are in progress to identify sources of tolerance. Screening for drought tolerance Wild accessions were screened in the drought prone area at RRS, Dapchari, Maharashtra, a region in West Central India experiencing high temperature and light intensity during summer months with soil moisture stress. Out of the 235 accessions screened, 23 potential accessions could be identified. In general, Mato Grosso and Rondonian accessions were superior for drought tolerance. Accessions RO 1769 and RO 2976 gave good juvenile yield under drought stress. These 23 accessions identified are now under a detailed evaluation at RRS, Dapchari along with 6 check clones viz., RRII 430, RRII 414, RRII 105, RRIM 600, RRII 208 and Tjir 1. After experiencing ten summer periods from 2008 – 2017, four wild accessions recorded girth higher than the proven drought tolerant clone RRIM 600. Accession MT 4856 recorded highest girth at 10th year under unirrigated condition at Dapchari and other promising accessions are MT 40 and MT 43. Polycross seeds were collected from this field trial to evaluate their growth and yield performance for selecting potential drought tolerant recombinants of wild and Wickham clones. Another accession RO 5363 was also identified as superior for yield in both the drought prone region at Sukma in Central India, and cold prone region at Nagrakatta in North East India. Screening for cold tolerance 64 wild accessions along with check clones are under evaluation in the cold prone areas in the Sub-Himalayan region at Nagrakatta, West Bengal, India. Accessions AC 3353, RO 2902, RO 3204, RO 2727, MT 915 and MT 900 recorded high girth and tappability compared to the check clone RRIM 600. Accessions AC 471, AC 4653, AC 3514 and MT 915 recorded relatively good yield. Germplasm Working Collection Field attrition over time can result in random loss of germplasm accessions, including the potentially useful ones. Hence the large number of accessions identified at different stages during the characterization and evaluation for yield/test tap yield, laticifers, disease and drought tolerance traits, are being established in a separate working collection to ensure better maintenance and easier access to the promising selections of wild germplasm for various uses. A total of 119 wild accessions have been established in the Germplasm Working Collection . Utilization Incorporation of the wild germplasm accessions in the breeding pool in India commenced in 1990 . Thirteen promising W x A hybrids evolved from the early crossings are now in the

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pipeline and are under final evaluation in Farmer Participatory trials across 10 locations in the traditional and non-traditional regions. With an aim to develop new generation high-yielding clones through transgressive introgression, hybridizations were carried out during 2011-14 using high-yielding Wickham clones (RRII 105, RRII 414, RRII 429 and RRII 430) as female parents and superior Wickham x Amazonian hybrids (95/10, 95/34 and 95/274) as male parents. Presently, 353 hybrid seedlings from 14703 hand pollinations are being evaluated in seedling nurseries. Based on mean test tap yield over two years, 22% of hybrids were selected for the next stage of evaluation under clonal nursery trials. Details are given in Table 2. Hybridisation was continued in 2017 in the breeding orchard at RRII, Kottayam, using selected parental combinations. Hybrid progenies (100 nos.) from 2017 hybridization programme were planted in seedling nursery for further evaluation.

Table 2: Details of nursery selections from W x (WxA)hybrids

Year

2012 2013 2014 Total

No. of full sibs test tapped 92 139 54 285 No. of full sibs selected 14 30 19 63 Percent recovery of selections

15 22 35 22

Since 2009, 13 wild accessions including one with high tolerance to Oidium, have been used in five W x A hybridization programmes. A total of 275 progenies are under evaluation now. Hybrid progenies resulting from the HP 2009 programmes at CES and RRS Padiyur are under evaluation for their early growth performance. At Padiyoor, 14 hybrids of two parental combinations RRII 105 x AC 675 and RRII 105 x RO 368, were found to be promising on the basis of test tap yield and were multiplied for further evaluation. At CES, 5 potential selections were identified from crosses involving 3 wild accessions and 6 cultivated Wickham clones. In the 2013 HP programme involving 2 potential high yielders and RRII 105, 8 progenies with promising test tap yield were selected for further evaluation in FETs, while another 11 hybrids were selected for clonal nursery evaluation. Among the 2014 HP progenies involving 4 wild accessions with RRII 105, 6 with promising test tap yield were selected for further evaluation in FETs. All these belonged to the cross combination AC 4833 x RRII 105. Eleven progenies with good test tap yield were also selected for clonal nursery evaluation. In the 2016 HP, 3 wild accessions (relatively high yielders MT 4788 and RO 4599, and Oidium tolerant RO 2871) crossed with RRII 105 resulted in 100 progenies which were planted in the seedling nursery. The family of the Oidium tolerant cross showed the highest girth. In an attempt to evaluate the potential of open pollinated progenies of the wild germplasm, 482 seeds from two FETs comprising potentially high yielding wild germplasm accessions interspersed with Wickham control clones, are being raised. III. General Hevea breeding programmes in India

1. Hybridization and clonal selection While newer hybridization programmes were taken up during the year, hybrids evolved from earlier HPs were evaluated over the long term in 15 small scale trials. Present hybridization programmes are directed towards disease resistance breeding and incorporation of wild Amazonian germplasm in the breeding pool for specific purposes.

27

2. Ortet selection Ortet selection was restricted to clonal nursery evaluation of ortets selected from the Andamans and large scale evaluation of ortet clones derived from ortets selected from large estates and small holdings. In clonal nursery trial (2012) of Andaman ortets at Chethackal, ortets selected from the oldest surviving seedling populations (source GG1 and GG2 seedlings of PBIG, Malaysia) located at south Andamans, were assessed in the 4th year of planting. Yield of ortets ranged from 21 g/t/15t (And-Or 103/8) to 111 g/t/15t (And-Or 104/10) compared to check clone RRII 105 (75 g/t/10t) and RRII 430 (150 g/t/10t). Girth of best ortet (104/10) was 22 cm compared to RRII 105 (18 cm) and RRII 430 (23 cm). 3. Large scale evaluation (LST) In the two LSTs of RRII 400 series clones at CES (LST I and LST II), significant clonal variation for yield under stimulation in the 15th and 16th year was detected. In LST I, RRII 417 (79g/t/t) out yielded RRII 105 (69 g/t/t) under stimulation. In LST II, PB 330 (91 g/t/t) was superior to RRII 105.RRII 430 also performed well with >70 g/t/t in both years. In the 1994 trial, where 12 indigenous and exotic clones are under evaluation, average yield over 13 years of tapping showed that clones 86/44 (73 g/t/t), RRIM 722 (70 g/t/t) and 86/120 (70 g/t/t) had yield performance comparable with that of RRII 105 (71 g/t/t). In a study on response of above clones to stimulation, clones RRIM 712 (133 g/t/t), 86/120 (119 g/t/t), and RRIM 722(118 g/t/t) exhibited high yield under stimulation. 4. On-farm evaluation (OFT) To evaluate the performance of pipeline clones in the South Karnataka region, an on-farm trial was laid out at KFDC, Karnataka, with fourteen high-yielding pipeline clones and seven control clones. The above pipeline clones are under evaluation under PCE Phases I (2008), II (2010) and III (2012), in various locations. The clones in the above on-farm trial were evaluated for growth performance in the third year of planting. Six pipeline clones showed better growth compared to check clone RRII 105. Pipeline clone P 102 had maximum girth followed by RRII 414 and RRII 430. 6. Polycross progeny evaluation Evaluation of progenies of nine prepotent clones over a period of 24 years for growth, yield and secondary attributes in two field trials planted in 1993 was completed. Timber volume was estimated. Wood samples were collected for laboratory studies on quality parameters. The 35 selections made from these progenies were classified as dual purpose latex-timber clones and timber –latex clones as well as latex clones and timber clones. Fifteen clones exhibited stability in performance over 12 years of tapping . The most promising 12 selections in the pipeline, out yielded the check clone by over 20%. Response to stimulation was studied in the 13th to 15th years of tapping. The selections in the pipeline are now under participatory evaluation trials in various locations.

Performance of polyclonal seedlings and check clone RRII 105 was assessed in 5th year of tapping. Mean yield of polyclonal seedlings was 59.6 g/t/t compared to check clone, RRII 105 (42 g/t/t). Six polyclonal seedlings had more than 120 g/t/t and 24 had yield more than 90 g/t/t with potential for further selection. Polyclonal seedlings were also better in terms of girth. Girth of polyclonal seedlings (Fig. 1) ranged from 45-112 cm (mean girth, 73.6 cm) as compared to RRII 105 (mean girth, 58.5 cm).

28

Fig 1 Distribution pattern of polycross progenies of Nettana origin based on girth 12 yr planting (n=172) (x-axis, girth in cm) Under the experiment on estimation of out-crossing rate in polyclonal seed garden of rubber at Kanyakumari, second set of progenies were evaluated for growth and yield performance at CES, Chethackal. Test-tap yield of progenies ranged from 3 to 258 g/t/10t in the third year after planting. Twenty four progenies had more than 90 g/t/t, nine progenies with more than 150 g/t/10t and three progenies were out-standing with more than 200 g/t/10t. Overall, forty nine progenies performed well above the population mean of 58.4 g/t/10t. The above population opened tremendous scope for selection of considerable number of progenies for advancement to further stages of clonal selection. 6. Investigations on Genotype x Environment interactions. Growth and yield of pipeline clones was assessed under cold and drought conditions at Agartala and Dapchari, respectively. Under drought conditions at Dapchari, P 102 (26 cm) and P 26 (22 cm) performed better than RRII 208. Fourteen pipeline clones were found better than RRII 430. Regarding test tap yield, P 20 topped yield along with RRII 430 and (50 g/t/15 t) and eleven clones performed better than RRII 105. At Agartala, under cold condition, pipeline clones P 107, P 102, RRII 429 and P 21 (34 cm) showed maximum girth while 10 clones were better than RRII 105. Regarding test tap yield, P 21 and RRII 430 (138 g/t/15 t) topped with maximum yield. The above observations indicated local adaptive nature of pipeline clones under contrasting environs. In the G x E interaction studies initiated in 1996 in popular clones across five locations, collection of yield data was concluded from three LSTs in Kanyakumari, Agartala and Nagrakata before project completion. Among the other two locations, in the LST at RRS, Padiyur, where commencement of tapping was delayed, yield data collection continued in the renewed bark. Yield of the RRII 400 series clones except RRII 429 and RRII 414, showed a rising trend and significantly superior yield than RRII 105 in the 10th year of tapping. Clone RRII 430 showed highest yield (65 g/t/t) followed by RRII 417 and RRII 422 (64-60 g/t/t) as compared to RRII 105 (45 g/t/t). GEI analysis for long term yield across four diverse locations was attempted using different models. In the LST at Odisha, in drought prone Central India where the trial was affected by cyclone, long term data on growth, yield, DRC and structural parameters was summarised . Clones RRII 429, RRII 430 and RRII 422 showed significantly superior yield than the check clone RRIM 600.

29

The adaptive plasticity of RRII 430 and RRII 422 was reconfirmed from the GEI analysis based on temporal yield variation from the region. Clone RRII 429 was specifically adapted for the region and RRII 414 showed high GEI (Fig. Bot 3). Based on the findings, seven clones including RRII 417, RRII 203, PB 217 and RRIC 100 could be suggested for further large-scale evaluation in Eastern India region.

Fig Bot 3. GGE biplot of ‘clone x year’ interaction for dry rubber yield among 12 clones. Clones at the right side the line perpendicular to the average environment axis are highly suitable for growing in the Eastern Indian region. Genotype ranks fall as they fall back from the ideal genotype

7. Participatory evaluation of rubber clones in the pipeline

Source Bush Nurseries of Pipeline clones At the Central Experiment Station, 267 pipeline clones were maintained in 16 source bush nurseries. Fourteen clones selected from clonal nursery evaluations of ortets and half sib selections were added to the pipeline, making the total number of pipeline clones 281. A total of 90pipeline clones are under evaluation in six Central Large scale Trials at the Central Experiment Station and RRII Farms and 39 satellite On farm Trials in farmers’ fields and large estates under five phases of Farmer Participatory Clone Evaluation. Mature trials under Phase 1 (2008) In the Central LSTs which were in the first year of tapping, in terms of girth in LST I, clones P 21, P61and P 74 along with the checks, RRII 414 and RRII 430 were vigorous. Clones, P021, P066 and P073 gave higher yield than RRII 414, the highest yielding check (with 60 gtt).Seven clones were on par with RRII 414. In LST II, Clones RRII 414, P 44 , P 26 and RRII 430 were vigorous in terms of girth. Three clones had higher DRC than RRII 105. P044 was the highest yielder. Seven clones were comparable to RRII 414, the highest yielder among the check clones. In the OFT at Vithura, among the pipeline clones, maximum girth was recorded in P 61 (65 cm) followed by P 21 (63 cm), P 84 (63 cm), P 88 (62 cm) and P 68 (62 cm). Minimum girth was recorded in P 60 (57 cm). In the OFT at Kanjirappally, P 26 recorded 89.4% tappability. Even though the clones generally showed poor growth due to local conditions, seven pipeline clones showed better growth compared to RRII 430, but none were superior to RRII 414. In the OFT at PCK, Athirapilly estate, trees were assessed for tapping during 3rd year of tapping with d/3 system of tapping. Maximum mean yield was recorded in P 067 (77 g/t/t) when compared to check clone RRII 430 (49 g/t/t). Pipeline clone P 074 (36 %) followed by P084 (35 %) recorded

Yr1

Yr2

Yr4

Yr3

RRII

RRII

RRII RRIC RRII

RRII

RRII RRII RRII

PC1 –

PC

2 –

- - - 0 2 4 6

--

0

24

6

Average environment

Ideal genotype

8

RRII RRIM

PB21

30

maximum DRC. In the OFT at Calicut Estate, during 3rd year of tapping, maximum yield (g/t/t) was recorded by RRII 414 (36 g/t/t) followed by P 067 (28 g/t/t). PCE trials under immature Phases 2 to 5 The Phase 2 trials laid out in 2010 were nearing tappability while the trials laid out in 2012, 2014 and 2016 in subsequent phases were at various stages of immaturity. 8. Breeding for drought tolerance In the SST 1998 A, four selections with high un-stimulated yield were subjected to second round of ethephon stimulation. Ortets 272 and 58 showed better response than RRII 105 with 50 % improvement in yield. Among the ortets selected from Dapchari, in drought prone West Central India, Dap 111 responded better to stimulation with 40 % yield improvement. In SST 1999 A, analysis of average yield over nine years showed maximum yield in 94/50 (70 g/t/t) followed by 94/23 and 94/44 (69 g/t/t) and 94/101 (64 g/t/t). The clones were assessed for response to stimulation. Maximum yield after stimulation was recorded in 94/50 (108 g/t/t) followed by 94/44 (108 g/t/t). In an attempt to develop drought tolerant clones for the non-traditional area, the drought tolerance capacity of the selected progenies, developed through hybridization between high yielding clone (RRII 105; female parent) and a drought tolerant clone (PB 280; male parent) and the reciprocal crossing (PB 280 x RRII 105), were evaluated in a clonal nursery trial in the drought prone area (RRS, Dapchari). Forty trial clones along with nine control clones were planted in the trial. Sixteen clones in terms of girth and five clones in terms of test tap yield were found superior to the check clones RRII 422 and RRII 417. Selection 69 showed superior growth and yield performance. Growth and yield performance of selections was monitored in 13th year of tapping among 15 hybrids and check clones in 1998 A SST at CES. Selections, which are also under evaluation in Phase 5 of PCE viz. 93/214 (69 g/t/t) and 93/216 (57 g/t/t), maintained superior yield trend compared to RRII 105 (45 g/t/t). 9. Breeding for disease resistance Hybrid progenies developed under breeding for disease resistance programme showed good test tap yield up to 63 g/t/10t in Fx 516 x RRII 414 and 34 g/t/10t in RRII 105 x Fx 516. First ever set of hybrids generated through hybridization between RRII 400 series and Oidium tolerant germplasm accession have been recovered and progenies were planted in seedling nursery for further evaluation. Second set of Fx 516 OPs were assessed for yield and screened for tolerance to Corynespora. It was possible to recover very high-yielding half-sib selections from Fx 516 with test-tap yield up to 93 g/t/15t. The Fx 516 half-sibs as well as hybrids were screened for disease resistance in collaboration with Protection Division. Half-sibs and hybrids, which are highly tolerant to Corynespora by leaf bio-assay, were also identified. 10. Studies on propagation A study aimed at developing drought tolerant rootstocks for the non-traditional area by evaluating the drought tolerance capacity of the seedlings from non-traditional areas compared to seedlings from traditional areas, was carried out in a nursery trial at RRS, Dapchari. Seeds were collected from three drought prone non-traditional rubber growing areas namely Maharashtra (RRS, Dapchari), Orissa (RRS, Dhenkanal) and Karnataka (HBSS, Nettana) and from traditional areas namely Kerala (CES, Chethackal) and Tamil Nadu (HBSS, Paraliar). Assorted seeds as well as seeds from polyclonal seed gardens and drought tolerant clone RRIM 600 and drought susceptible clone RRII 105 were collected from each of above locations. Seeds were also collected from drought tolerant clone RRII 203 located at Orissa. The seedlings were maintained under rain-fed condition at Dapchari. Bud-grafting of those seedlings which survived drought condition at Dapchari was carried out using drought susceptible clone RRII 105 as scion. Bud-

31

grafting of the unselected seedlings at Central Nursery Karikkattoor was also done using RRII 105 as scion. Field planting of plants which survived drought as well as the control plants was carried out at RRS, Dapchari for further evaluation. 11. Region-specific clone recommendation The formulation of region specific clone recommendations based on agroclimatic zoning was a long felt need of the rubber plantation sector, so as to enable maximisation of rubber yield from each region and thereby improve rubber production per se. Based on results of 29 field evaluations of rubber clones in the traditional region as well as studies on disease reaction, genotype x environment reaction studies and yield from several large scale and on farm evaluations of these clones in Tripura , Assam and Northern part of West Bengal in North East India region specificity in performance of the clones was determined. A new region specific advisory of clones suited to each of the nine agro climatically distinct zones in the traditional and non traditional regions of rubber cultivation India was formulated and approved in the reporting year for communicating to the rubber plantation sector. Region specific clone advisory for the rubber growing regions of India

* RRII 429 has proved to be high yielding and can be considered for planting in locations as recommended above in the traditional tract subject to effective management of pink disease. Clone RRII 430 is not suitable for high altitude regions and NE India where high severity of Oidium infection is experienced. @ Recommended plant protection measures are advised in all rubber growing regions. # The system of tapping recommended for the above high yielding clones is 1/2Sd3. Clone RRII 105 can also be tapped at lower frequencies as recommended by the Rubber Board.

Region States/ Districts Clones recommended * @ #

North East India Tripura, Assam, Northern West Bengal, Meghalaya

RRII 208, RRII 429, RRIM 600

East and West Central India

Maharashtra, Odisha RRII 429,RRII 208, RRII 430, RRIM 600

South-West Karnataka and Goa

Goa and Dakshin Kannada

RRII 430, RRII 414, GT 1, RRII 203.

North Kerala Kozhikode,Malappuram, Kannur, Kasargode

RRII 430, RRII 417, RRII 105

North Central Kerala Trichur Palghat. RRII 417, RRII 429, RRII 430

Central Kerala Ernakulam, Kottayam, Pathanamthitta, Alleppey

RRII 430, RRII 414, RRII 417

South Kerala Kollam, Trivandrum RRII 422,RRII 430, RRII 417

Tropical high altitude regions

Wynad, Idukki, Kulathupuzha, Vithura.

RRII 422, RRII 429, RRII 417

South Tamil Nadu Kanyakumari RRII 430, RRII 105, RRII 429

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6. REPORT ON HEVEA BREEDING ACTIVITIES IN MYANMAR

I. Multilateral Clone Exchange Programme Myanmar has so far received 21 clones from five countries under the multilateral clone exchange program. Of these, 16 clones from Philippines, India , Thailand and Ghana (CIRAD) have been established in source bush nurseries . A total of 1024 budwood points are thus maintained. Five clones received from Sri Lanka have been multiplied along with additional budding done for the clones from Ghana and 749 plants are to be planted in SBNs this year. Details of the MCE clones are as follows:

Sl. No.

Country Date received

Clone Budwood received (m)

SBN plants

already planted

Plants to be

planted

no. of stocks being

budding

1 Philippine 24.8.2016 USM 1 21 41 - -

Total 1 21 41 - -

2 India 7.9.2016 RRII 414 5 153 - 601

RRII 417 5 58 - 152

RRII 422 5 153 - 617

RRII 429 5 153 - 1203

RRII 430 5 119 - 400

Total 5 25 636 - 2973

3 Thailand 13.6.2017 RRIT 226 62 - 250

RRIT 3604 89 - 140

RRIT 408 58 - 185

RRIT 3904 78 - 260

RRIT 251 44 - 140

Total 331 - 975

4 Ghana 17.8.2017 FDR 5788 6 5 90 -

CDC 321 6 6 100 -

FDR 5665 5 2 45 -

FDR 4575 6 1 40 -

PMB 1 5 2 30 -

Total 28 16 305 -

5 Sri Lanka 24.1.2018 RRISL 2001 5 - 100 -

RRISL 211 5 - 70 -

RRISL 219 5 - 85 -

RRISL 208 5 - 82 -

RRISL 203 5 - 107 -

Total 25 - 444 -

Total 21 99 1024 749 3948

II. General Hevea breeding programmes in Myanmar

The progress of Hevea breeding in Myanmar during 2018-19 can be summarised as follows:

• Multiplication nursery was established including 21 exchange clones and some local developed hybrids.

• Hand pollination was carried out but failed to get good seeds because of the heavy rain.

• Germplasm collection plot was set up with the selected local developed hybrids and some introduced clones from Malaysia, Indonesia, China, etc., according to FAO project.

• Breeding plot was established for hand pollination.

33

• The data was recorded from the small scale clone trails with the selected local developed clones already established in 2005-06 and 2013-14.

• Budded stumps of the local clones selected from SSCT established in 2005-06 were sent to different locations to set up the clone trail and multiplication bud-wood nursery.

7. REPORT ON HEVEA BREEDING ACTIVITIES IN RRISL, SRI LANKA

I. Multilateral Clone Exchange Programme During the reporting year, sent Import Permit for clones from Ghana and Cote d’ Ivoire.

Table : Details of clones received and their bud grafting success

Country Clones Number of bud grafted

Number of success

India RRII 422 12 10

RRII 429 25 24

RRII 417 5 4

RRII 414 15 6

RRII 430 6 6

ARCPC 24/4 26 2

ARCPC 6/22 43 37

RRIT 3604 9 9

RRIT 251 23 23

RRIT 408 18 16

RRIT 3904 23 19

RRIT 226 18 16

II. Conservation and evaluation of the IRRDB germplasm (GPB/GP/85/2)

Multiplication and evaluation of the genotypes collection of Hevea obtained from 1981 IRRDB expedition to the Amazon Multiplication/ Establishment and scientific evaluation of the Hevea germplasm collection was started with the aim of enhancement of productivity through genetic improvement and management of genetic resources of Hevea. Around 1400 accessions were planted at Nivitigalakele Substation and some accessions are ready to establish in bud wood nurseries at Neuchatle estate. Thousand eight hundred and thirty number of accessions were selected for this year multiplication and establishment. All these accessions were preliminary characterized under different categories such as early and late wintering, high girth, dwarf, tall, smooth bark etc. Then they were pollarded and established the seedling nursery for bud grafting. Completed the planting in Nivitigalakele substation. Testing of promising clones for sub optimal conditions Evaluation of adaptability and performance of new promising clones in nontraditional rubber growing areas (sub optimal conditions) is aimed with this project. All trails are conducted as in RRI/smallholder collaborative manner. III. General breeding programmes in RRISL Molecular applications 1. Screening of new genotypes developed using the germplasm of Hevea obtained from

1981 IRRDB expedition to the Amazon (RRI/ECT collaborative trial 2011-Yatadola estate) Ten genotypes selected from 1995 hybridization programme where non-Wickham clone GPS 1 was used as male parent, were characterized. Fifteen Microsatellite molecular markers were screened and three distinct groups were developed. Other parameters such

34

as girth, bark thickness, bole height, branching pattern and canopy density were also used in genotype screening.

2. Screening of selected Hevea brasiliensis L. genotypes from 2005 HP progeny for Corynespora leaf fall disease (CLF) resistance using Microsatellite Molecular markers Thirty Microsatellite molecular markers were screened with parents, grandparents and moderately susceptible control clones and found polymorphism in four primers and used to assess progeny along with six clones (RRIC 100, RRIC 103, RRIC 52, PB86, RRISL 201 and RRISL 208. In the field screening it has observed that the clone RRIC 100 and PB 86 were resistant, RRISL 201and RRISL 208 were moderate resistant and RRIC 103 and RRISL 201 were susceptible to CLFD. Pairwise genetic distance matrix was developed.

Analysis of antioxidant gene expression in tapping panel dryness (TPD) affected rubber tree (Hevea brasiliensis Muell. Arg.) and the effect of exogenous application of Ascorbic acid on alleviating TPD This was started with the objectives of studying the role of antioxidant genes on TPD and the effect of exogenous application of antioxidants on TPD affected rubber trees. Using quantitative real time PCR, the expression pattern of antioxidant genes will be analyzed in TPD affected and healthy rubber trees to identify which antioxidant genes express deferentially under TPD condition and to understand their involvement in TPD since this information will be vital for designing a treatment or future molecular remediation, such as generating transgenic rubber plant overexpressing antioxidant genes, to cope with TPD and in turn to minimize the economic losses caused by TPD. According to the results, HbSOD gene is down regulated in all the TPD affected rubber clones whereas HbGPX and HbCAT genes are up regulated in all the TPD affected clones. Based on the result, it seems that HbSOD gene may play a significant role in TPD. TPD affected laticifer tissues might run short of antioxidants mainly HbSOD antioxidant enzyme making the clone more likely to get TPD. Ascorbic acid application of TPD affected trees It is worthwhile to study the effect of exogenous application of antioxidants such as ascorbic acid to compensate this shortage of antioxidants and the potential of ascorbic acid to recover TPD affected clones (third objective of above research) This experiment is started at two sites as Eladuwa and Payagala with four clones, three levels of Ascorbic Acid concentrations (5mM,10mM,15mM) with control ( water with surfactant). Each treatment got five replicates. Around fifty days after starting application started record the recovery percentage while continuing the application. Data were collected in three cycles and done the statistical analysis. It has observed that considerable recovery percentage from 5 percent to 20 percent compared to control in all clones in both sites. However, due to a severe cyclone affect more number of trees were damaged and was shifted to Payagala estate. Evaluation of previous hand pollinated (HP) progenies Small Scale Clone Trials Nineteen small scale clone trials which maintained and monitored during the year However, due to heavy rainfall received throughout the year, the yield data collection in most of the trials could not be possible.

35

8.REPORT ON HEVEA BREEDING ACTIVITIES IN RRIT, THAILAND Ratchanee Rattanawong I. IRRDB Multilateral Exchange Clones In Thailand Since 2014, Thailand imported rubber budwood of 5 clones of from India, 5 clones China, 5

clones from Cambodia, 5 clones from Sri Lanka, 2 clones from Myanmar, 5 clones from Vietnam, 5 clones from Cote d’Ivoire and 5 Clones from Ghana. For these reason, Five RRIT’s clones (RRIT 226, RRIT 251, RRIT 408, RRIT3604 and RRIT3904) budwood already sent to India in October, 2014, to Philippines on 29 September, 2015, to CRRI on November, 2015, to Sri Lanka, Cote d’Ivorie, China in September and October, 2016, respectively.

In 2017, Five RRIT’s clones already sent to Vietnam, Cote d’Ivoire and Ghana on September, October and November 2017 respectively.In 2018, new Import permit for the remain member countries already released. Indonesia is the first countries that start going in 2018. Five RRIT’s clones already export to Indonesia on July, 2018.

Table 2 The status of IRRDB Multilateral Clone Exchange program in Thailand

Country Import/Export

Date Exchange Clones

India Import October, 2014 RRII 422, RRII 429, RRII 417, RRII 430, RRII 414

Export November,2014 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Cambodia Import October, 2015 FDR 5788, CDC 312, PMB 1, FDR 5240, FDR 4575

Export November, 2015 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

China

Import September, 2015 Reyan 7-20-59, CATAS 7-33-97, Hongshan 6-7-15, Zhanshi 8-67-3, Baoting 936.

Export September, 2015 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Philippines Import On going

Export November, 2015 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Sri Lanka Import November, 2015 RRIC102, RRISL203, RRISL2001, RRISL2003, RRISL 2005

Export November, 2015 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Myanmar Import August. 2016 ARCPC 6 (22) , ARCPC 2 (4)

Export August. 2016 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Vietnam Import September, 2017

RRIV 1, RRIV 5, RRIV 106, RRIV 107, RRIV 114

Export September, 2017

RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Cote Import October, 2017 IRCA 41, IRCA 230, IRCA 331,IRCA 317,IRCA 804

d’Ivoire Export July, 2016 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Ghana Import December, 2017 CDC 312, PMB 1, FDR 5240, FDR 4575, FDR 5788

Export November, 2017 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

Indonesia Import July, 2018 IRR 5, IRR 104, IRR 119

Export July, 2018 RRIT226, RRIT251, RRIT 408, RRIT3604, RRIT 3904

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II. Status of the 1981 IRRDB Wild Hevea germplasm in Thailand

Cooperation between IRRDB and Brazilian government expedition was carried out in 1981 to collect wild Hevea germplasm in Amazon rain forest, Brazil. As a member of IRRDB, Rubber Research Institute of Thailand (RRIT) received 6,069 germplasm from Malaysia germplasm center during 1985-1989. However, 3,085 wild germplasm are survived and conserved as budwood nursery in 3 locations as Chachengsoa Rubber Research Center, Nongkhai Rubber Research Center and Songkhla Agricultural Research and Development Center representative of East, Northeast and South region of Thailand, respectively. The characters and performance of the germplasm are continue evaluated. Moreover, 2,984 Hevea wild accessions also conserved as genepool garden with 5 trees per genotype in Traditional planting area (Surat Thani, Ranong, Phuket and Krabi province) and Non-traditional area (Nongkhai and Chachengsao province). Agronomic characters and growth vigor of the germplasm were evaluated. The genetic vigor of the germplasm was evaluated using girth measurement at the height of 170 cm above ground.According to the growth vigor information, in Non-traditional area, Mato Grosso germplasm had better average girth measurement than Rondonia and Acre germplasm. Whilst in traditional area, Acre germplasm showed the highest of girth. Latex yield potential also has been evaluated. Most of germplasm was opened for tapping using 1/2s d/2 tapping system. The various average yield of each district accession were investigated. Latex yield of Mato Grosso germplasm have been better than Acre and Rondonia germplasm. Otherwise, ortet clones in Phuket and Chachoengsoa genepool garden and Acre germplasm gave the highest average latex yield. From the result of the evaluation, the 1981 wild germplasm showed the variation of growth vigor and latex yield. Most of the genotype which expressed high girth but low yield.

The result of 1981 germplasm evaluation in genepool garden also showed large variation of growth vigor and latex yield potential. However, most of the genotype showed high girth measurement, the latex yield was generally low. From these performances, it is quite difficult to select and improve any high latex yield performance from the 1981 wild germplasm. But it is possible to use the 1981 wild germplasm as parents crossed with high yielding Wickham clones for improve high yielding clones along with good growth in breeding program. Consequently, elite of 1981 wild germplasm has been selected and used as parents to enhance the genetic base of breeding program.

The utilization of 1981 Hevea germplasm in RRIT breeding program According to the evaluation for girth and latex yield potential in the 1981 wild germplasm genepool garden in tradition and non-traditional planting area of Thailand. It is very unlikely to select the high yield performance of the 1981 wild germplasm used in breeding program for high latex yield clones. However, the progenies of Wickham and Wild germplasm hybridization are expected to show very low latex yield, it is possible to use these germplasm for genetic base enhancement. Promising genotypes of the 1981 wild germplasm was selected and used in breeding program, crossed with high yielding Wickham clones. Since 1992, Wickham x Amazonian (W x Am) crosses were attempted. Four hundred and fifteen crossing and 7,388 progenies were reported since 1992 to 2009. However, 6,124 progenies were tapped and evaluated, 589 progenies were selected and planted in Small Scale Clone Trial, consequently. In 2006, the highest number WxAm crossing (65 crossing) were operated, 2,135 progenies have been planted and evaluated in Seedling Evaluation Trial (SET). In the SET, the progenies were tapped at 2 or 3 years, depend on the girth measurement. Ten day tapping for 3 cycles using 1/2s d/3 tapping system were applied. After 3 cycles tapping test (30 tapping days), 2.5 % stimulant was applied using the same tapping system. From the SET, 318 progenies were selected and transferred to the small

37

scale clone trial in the non-traditional rubber planting area such as Chachengsao and Nongkhai province and 561 progenies were selected and transferred to the small scale clone trial in the non-traditional rubber planting area such as Chanthaburi and Krabi province in 2002 to 2013.

Table 1 W x Am progenies at Chanthaburi Agricultural Engineering Research Center.

Clone Girth (cm.)

% G/T/T %

15.5Y R600 Y1 Y2 Y3 Y4 Y5 Y6 Y7 Y8 AV R600

OP-CH-38-1695 67.6 114 43.2 44.8 44.5 64.5 83.7 115.4 119 78.4 74.2 252

OP-CH-38-2258 85.4 145 24.6 39.6 34.8 57 81.5 105.9 102.8 83.1 66.2 225

OP-CH-38-2345 74.5 126 27.8 39.2 34.7 63.5 79.4 78.7 55.7 48.3 53.4 182

OP-CH-38-2021 55.5 94 34.3 52.6 46.3 53.8 65.1 85.1 51.8 33.5 52.8 180

OP-CH-38-2043 71.4 121 31.5 38.8 38.2 54.8 66.7 77.1 57.7 46.7 51.4 175

OP-CH-38-2118 66.1 112 22 36 36.2 54.2 65.7 73.1 66 53.3 50.8 173

OP-CH-38-1621 67.5 114 34.5 43.4 38.3 49.9 58.5 76.8 61.8 35.4 49.8 169

OP-CH-38-1691 61.2 104 26.8 34.1 34 43.4 60.5 59.3 63.1 43.5 45.6 155

BZ-CH-38-510 48.4 82 29.5 26.5 28.9 36.4 58.1 75.1 58.8 50 45.4 154

BZ-CH-38-467 56.8 96 21 32.6 35.2 47.4 55.9 67.5 49.4 31.4 42.6 145

BZ-CH-39-534 89.6 152 14.3 22.3 27.8 40.9 49.8 34.2 47.6 49.2 35.8 122

BZ-CH-39-813 78.8 133 13.4 23.6 26.3 38.5 40.5 49.9 41.9 39.9 34.3 117

BZ-CH-39-810 69.9 118 17.8 23.4 27.3 43.8 44.4 32.2 30.2 33.3 31.5 107

BZ-CH-39-938 73.3 124 17.1 22.8 25.1 33.6 35.2 42.9 37.7 37.7 31.5 107

BPM24 62.1 105 22.5 33.7 27.9 37 47.2 58.3 48.2 31.3 38.2 130

PB260 55.8 94 18.3 23.8 26.8 32.3 38 53.3 43.7 28.6 33.1 113

RRIM600 59.1 100 18.8 23.5 25.6 33.2 32.3 34.8 33.4 33.4 29.4 100

III. Hevea breeding in Thailand

Rubber breeding in Thailand started from 1933 until now by the conventional rubber breeding program to develop rubber clones using many methods such as importing the rubber clones from abroad, hand pollination and rubber clonal selection. Because of rubber planting area in Thailand expanded to northeast and north of Thailand very rapidly, rubber planting has increased from 1.92 million hectare in 1997 to 3.67 million hectare in 2016. Low fertility of soil and unsuitable climate condition is the limited factor for planting rubber in new planting area. Thus, improving new rubber clones suitable for new planting area through breeding program started since 1992. Moreover, Tissue culture and biotechnology have been applied for rubber clonal improvement. The objectives of rubber breeding included high latex yielding, high timber yielding, tolerant to rubber diseases and good adaptation for the specific environment. Conventional breeding in marginal area has been carried out following the standard procedure. The procedure start from hand pollination, selection in the nursery, small scale clone trial and large scale trial. 1. Hand pollination: Since 1992 – 2017, number of 3,035 - 55,660 flowers were pollinated at Chachoengsao Rubber Research Center by Kannikar Teerawatanasuk and 351 flowers were pollinated for resistance to Phytophthora leaf fall at Surat Thani Agricultural

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Research and Development Center in 2017. Achievement of Fruit set rates varies from 1.09% to 8.87%, with average 4.43%.

2．Selection 2.1 Seedling Evaluation Trial (SET): The hybrid seeds obtained from hand pollination are first planted in polybags and at the two-whorl leaf stage, then transferred to the field using high density 2x2 m. The preliminary recording on vigor as expressed by diameter and girth are made at 10 cm high level every six months for two years. After the second year, the seedlings are test-tapped following s/2 d/3 tapping system. Each genotype evaluated based on ten–day-tapping system. In 2016, 135 genotypes were selected, the progenies of RRIT 251 x RRIC 130, BPM 24 x RRIT 408, and BPM 24 x RRIT 251 expressed high latex yield.

2.2 Small Scale Clone Trial (SSCT): The selected clones from SET has been propagated and tested in small scale clone trials which are presented at RRIT experimental station at Chachoengsoa, Nongkhai and Surat thani Rubber Research Center. Simple and triple lattice design with two and three replications are used and each plot consisted of 6-8 trees. Clone RRIM 600 is usually used as control. Girth measurements are taken at 170 cm height, Early selection are applied in SSCT. Since 2008, 24 SSCT have been set up and 8 genotypes expressed higher latex than control significantly as RRI-CH-36-980, RRI-CH-36-1035, RRI-CH-36-187, RRI-CH-36-1286, RRI-CH-36-301, RRI-CH-36-387, RRI-CH-36-1457, RRI-CH-36-1515 were selected in 2016.

2.3 Large Scale Clone Trial (LSCT): LSCT are agricultural trials carried out in condition similar to that of the planters. Generally experimental design is randomized complete block with 3-4 replications. Evaluated based on latex yield and secondary characteristics are taken at regular interval. The selected clones could be recommended for large scale planting. In 2008, 230 genotypes were evaluated in 18 LSCT in different locations. New hybrid clones, RRI-CH-35-1396 were selected and recommended as class 1 clones in 2011 clonal recommendation, namely RRIT 408. In 2016, 164 genotypes were evaluated and 4 genotypes expressed higher latex than control significantly as RRI-CH-35-1886, RRI-CH-35-779, RRI-CH-35-1323 and RRI-CH-35-1316.

According to the result of rubber breeding projects, Rubber Research Institute of Thailand’s (RRIT’S) clonal recommendations have been revised every 4 years based on information of latex yield from clone trials in traditional and non-traditional rubber planting area. Six RRIT’s rubber clonal recommendation, in 1993, 1999, 2003, 2007, 2011 and 2016 have been issued in this phase. The newest, 2016 of RRIT’s Rubber clonal Recommendation has been improved. Different climate zone of planting areas used as main criteria for considering, consequently, divided into 2 zones ; 1) Traditional zone, rubber clones recommended for traditional rubber planting area, included most of planting area in 14 provinces of southern area and 3 provinces in eastern area which have rainfall greater than 1,600 mm./year. And 2) non-traditional zone, rubber clones recommended for new planting area, promising clones selected based on the results from clone evaluation trails in non-traditional planting area which rainfall less than 1,600 mm./year (5-6 months of dry period) Clones for each zone separated into 3 classes as follows: Class I: The class refers to clones which have been tested and grown widely. Information on

yield performance, disease susceptibility and other secondary characteristics are carefully gathered for at least 10 years (3 years tapping) from various large-scale trials. The class I clones can be planted throughout a plantation.

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Class II: The class refers to clones which express high yield and good growth performance, but limited some information on yields and other characteristics. It is recommended that these clones have not planted over 50% per plantation or planted with the clone in class III not over 50% per plantation.

Class III: The class refers to clones which express high yield and good growth performance,

but limited some information on yields and other characteristics. It is recommended that these clones have not planted over 50% per plantation under the RRIT’s advise.

In addition, based on latex and timber yield potential, each class consists of 3 groups:

Group 1 : Latex clones, are clones which have high latex yield. These clones suitable for latex production only. Group 2 : Latex timber clones, are clones which have high latex yield with good growth and

long straight boles. These clones suitable for latex and wood production Group 3 : Timber clones, are clones which have high rubber wood production. These clones

have vigorous growth of stem, good long straight boles and good branching behaviour. However, the latex yield generally lower than group 1 and 2. These clones suitable for forest plantation.

RRIT’s clonal recommendation, 2016 is as follows : I) Traditional planting area Class 1, clones can be planted throughout a plantation Group 1 : Latex clones : RRIT 251, RRIT 226, BPM 24 and RRIM 600 Group 2 : Latex Timber clones : PB 235 and PB 260 Group 3 : Timber clones : AVROS 2037 and BPM 1 Class 2, clones can be planted not over 50% per plantation or planted with the clone from class III not over 50% per plantation. RRIT 403, RRIT 406, RRIT 408, RRIT 3702, RRIT 3801 and RRIT 3802 Class 3, clones can be planted not over 50% per plantation under the RRIT’s advise. RRIT 3902, RRIT 3903, RRIT 3904, RRIT 3905, RRIT 3908, RRIT 3909, IRCA 825 and IRCA 871 II) Non-traditional planting area Class 1, clones can be planted throughout a plantation Group 1 : Latex clones : RRIT 408, RRIT 251, RRIT 226 and RRIM 600 Group 2 : Latex Timber clones : RRII 118 and PB 235 Group 3 : Timber clones : RRIT 402, and BPM 1 Class 2, clones can be planted not over 50% per plantation or planted with the clone from class III not over 50% per plantation. RRIT 411, RRIT 417, RRIT 3604, RRIT 3607, RRIT 3609, RRIT 3610, RRIT 3612, RRIT 3702, RRIT 3902, RRIT 3904 and RRIT 3906 Class 3, clones can be planted not over 50% per plantation under the RRIT’s advise. RRIT 3608, RRIT 3611, RRIT 3613, RRIT 3614, IRCA 825 and IRCA 871

9.REPORT ON HEVEA BREEDING ACTIVITIES IN RRIV, VIETNAM Tran Thanh

I. Status of Multilateral Clone Exchange As an IRRDB member, the Rubber Research Institute of Vietnam has participated in this

program in the hope that the exchanged clones could be used as planting materials for

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commercial cultivation and/or parental materials in hand pollination program to widen the genetic base for genetic improvement in the IRRDB member countries. All clones included in this exchange program are elite RRIV’s clones, which have been contributing to the increase in rubber production through activities of replanting and enlarging the areas under rubber cultivation throughout the country.

Currently, a total of 13 clones were successfully exchanged between the Rubber Research Institute of Vietnam and other two rubber research institutes including the Rubber Research Institute of India (08 clones) and Rubber Research Institute of Thailand (05 clones). In addition, the bilateral clone exchange (05 clones) between the Rubber Research Institute of Vietnam and CIRAD – Michelin is in progress.

II. Progress in evaluation and utilization of the IRRDB 1981 germplasm II.1. Ex situ conservation of Hevea germplasm Currently, about 3,500 accessions of Hevea brasiliensis have been collected and conserved

in Vietnam. The majority of this germplasm were derived from the IRRDB’81 germplasm collected in the Amazonian habitats of the genus. This collection has been considered as the key factor contributing to the improvement of rubber tree through breeding programmes.

In Vietnam, the ex situ conservation of Hevea germplasm was established in 1986. The

majority of this germplasm collection was derived from the IRRDB’81 collection expedition in the Amazon forests of Brazil which is the primary center of diversity of the crop and the source of wild rubber trees. In addition, this germplasm also included the collection of local Hevea accessions, the introduction of Hevea clones from other countries. The IRRDB’81 germplasm was introduced into Vietnam in the form of budwood in 1984.

On receipt of the budwood, each accession was first multiplied by bud grafting and then

planted in the field gene-bank for conservation in the form of source-bush garden, which was laid out in randomized complete block design in which each accession was represented in two replications of five trees with planting distance of 1.5 m x 1.2 m. The genetic resources of Hevea germplasm conserved in Vietnam are shown in Table 1. The source-bush garden is cut back every year to maintain the conservation and also to generate budwood for various evaluation trials. Several preliminary field evaluation trials for most of Hevea germplasm accessions have been being established in several representative locations to evaluate their agronomical and morphological characteristics.

Table 1. The genetic resources of Hevea germplasm conserved in Vietnam

Genetic sources Number of accessions

South America 3084 Acre (AC) 963 Mato Grosso (MT) 900 Rodonia (RO) 1116 Others 105 Africa 38 Ivory Coast 38 Asia 223 Vietnam 126 Malaysia 55 Indonesia 7 Sri Lanka 16

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China 5 Cambodia 1 India 8 Thailand 5

Total 3345

II.2. Evaluation and characterization of IRRDB’81 germplasm in Vietnam

Most of the Hevea germplasm have been tested in various clonal trials including arboreta and small cale clonal trials which were established in several representative locations. Observation and recordings were carried out on main agronomical and morphological characteristics such as growth, latex production and disease susceptibility for all trials. Further studies of anatomical characteristics of bark, physiological characteristics of latex properties were conducted on the small scale trial, STLK 85. Details of these clonal trials are given in Table 2.

In respect of latex production, IRRDB’81 collection exhibited very poor performance with

an average latex yield of around 16% of the level of currently developed Wickham clones after five years of tapping. Outstandingly, in the first three years of tapping, some IRRDB’81 accessions exhibited very good performance in latex yield with the production up to 154% of that of the control clone. These accessions have been being used as parents in Hevea breeding programmes. A large number of IRRDB’81 accessions showed very good growth performance, especially in the highlands area which was considered as a non-traditional rubber growing region in Vietnam. Superiority of these accessions will be of considerable value in advanced breeding programmes. Recently, IRRDB’81 collection has been considered as an important source for timber selection and rubber wood production. As the results show, several IRRDB’81 accessions could be considered as suitable clones for timber production purposes.

Regarding disease susceptibility, differences in the level of resistance to important diseases have been observed in IRRDB’81 collections. It seemed that Wickham and IRRDB’81 accessions showed no significant differences in susceptibility to pink disease, powdery mildew and Gloeosporium leaf diseases. In the view of latex physiological parameters, the sucrose content (SUC) and plugging index (PI) of IRRDB’81 accessions were significant higher than the Wickham clones, but the latter have higher contents of thiols (RSH) and inorganic phosphorus (Pi). In IRRDB’81 germplasm, significant positive correlations were found between latex yield and RSH/Pi while significant negative correlation was found between latex yield and PI. On anatomical features of virgin bark, the IRRDB’81 accessions were significant lower than the Wickham ones in most of anatomical parameters of virgin bark, especially in latex vessel rings. In addition, the properties related to viscosity and plasticity of raw rubber in IRRDB’81 accessions were also studied. In general, three properties of rubber including initial plasticity, plasticity retention index and Mooney Viscosity were similar to those of rubber obtained from domesticated clones, and technologically and commercially acceptable.

Table 2. Details of arboreta (SG) and small scale clonal trials (ST)

Trials Location (province)

Area (ha)

Spacing (m x m)

Tree x Replication No. of accessions

Remarks

SGLK84 Binh Duong 3,00 7 x 2,5 2 x 3 235 Closed SGAL85 Dong Nai 7,00 7 x 2,5 1 x 5 648 Closed SGLK85 Binh Duong 15,80 7 x 2,5 1 x 5 1536 Closed STLK85 Binh Duong 12,50 7 x 2,5 8 x 3 200 Closed STLK86 Binh Duong 12,50 7 x 2,5 208 Closed

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STLK87 Binh Duong 12,50 7 x 2,5 3 x 2 216 Closed SGLK91 Binh Duong 1,40 7 x 2,5 1 x 5 133 Closed SGLK94 Binh Duong 4,95 7 x 2,5 3 x 2 328 Closed SGLK95 Binh Duong 2,08 3 x 3 12 x 2 52 Closed SGLK96 Binh Duong 3,00 7 x 2,5 3 x 2 225 Closed SGYS97 Binh Duong 1,80 (4+16) 3 x 2 3 x 4 57 Closed SGCS97 Gia Lai 2,00 7 x 2,5 1 x 5 204 Closed SGPH98 Phu Tho 1,24 7 x 2,5 1 x 5 153 Closed SGCP00 Gia Lai 1,00 7 x 2,5 3 x 2 81 Closed SGLK03 Binh Duong 2,00 7 x 2,5 3 x 2 190 SGLK04 Binh Duong 2,00 7 x 2,5 3 x 2 190 SGLK05 Binh Duong 3,10 7 x 2,5 3 x 2 265 SGLK06 Binh Duong 2,00 7 x 2,5 3 x 2 196

II.3. Utilisation of IRRDB’81 germplasm

Regarding widening the genetic base for genetic improvement, several promising IRRDB’81 accessions have been included in hand pollination program in RRIV since 1997. Based on the agronomical and morphological traits as well as the genetic diversity analysis, recently, many attempts have been made to enlarge the genetic base of Hevea breeding materials by polycrossing among different genetic resources. In this way, many crosses between Wickham (W) and Amazonian (A) clones have been preferably made, and of which the progenies are in various phases of evaluation with the expectation that they could not only combine the good agronomical characteristics from the parents but also help to enhance the genetic variability in the population. As the results, from 1997 to 2017, a total of 5,355 W x IRRDB’81 and IRRDB’81 x W progenies were produced from 529 crosses. Generally, in comparison to other progeny populations, these progeny populations exhibited rather good girth but very poor latex productivity in early selection trials. However, some progenies had significantly higher latex yield than the control clone in both early selection and small scale clonal trials.

III. Hevea breeding programmes

III.1. Hand pollination

Since the Hevea breeding program was resumed in 1980, many attempts have been carried out to exploit the available Hevea genetic resources for the hand pollination programmes. The origins of parents of hand pollination gardens showed that priority has been given to diversification of parental clones and combinations (Table 3). Besides, new Hevea germplasm, especially the IRRDB’81 collection, has been included in hand pollination programmes based on their performances in evaluation trials.

Table 3. The origins of parental clones in hand pollinations gardens

Origin Hand pollination gardens

1980 1984 1990 1993 1996 1999 2002* 2006*

Wickham Indonesia 9 2 Malaysia 15 8 1 11 26 4 5 Sri Lanka 14 3 2 3 2 5 Vietnam - Old clones 21 4 3 - Advanced hybrids 6 9 8 19 61 30 Others 2 4 2 South American

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Wickham x Amazon 24 3 2 5 2 1 Amazon 17 4 1 2 Others 1 1 7 2 IRRDB’81 germplasm Acre 4 10 6 3 Mato Grosso 3 10 4 Rondonia 12 2 10 14 4

Total 59 67 39 27 30 89 84 38

(*) These hand pollinations gardens are being used.

III.2. The network of clonal trials In Hevea breeding programmes, recombination, evaluation and release of clones is laborious

and time consuming. Recombination breeding starts with the production of full-sib families using the hand pollination via crossing highly heterozygous parents which is a traditional technique used. Each cycle of hand pollination involves a large number of crosses to generate few hundred to several thousand new progenies, therefore the first stage evaluation of these progenies is carried out in early selection trial (EST), that helped to reduce the potential individuals to a manageable level, followed by a small scale clonal trial (SSCT) and a large scale clonal trial (LSCT) and/or promotion plot. A small scale and large scale clonal trials are very important intermediary trialing stages in which the selections from ESTs are evaluated in normal commercial scale spacing to further reduce the number of selections to much smaller number, which are subsequently worth passing forward to onfarm trials.

In Vietnam, rubber-growing areas are mainly in the southeast region, followed by the Highlands, Coastal region and the newly-developed areas in the northern region. Of these regions, the southeast region is a traditional area for rubber tree, however, diseases are factors hindering the performance of clones in this region. In contrast, the Highlands are considered as a non-traditional region which is characterized by lower temperature, drought, regularly stronger wind and higher number of mist days than the Southeast region. The Coastal region is characterized by many constraints such as very strong wind caused by typhoon and coldness which displayed adverse effects on the survival of rubber trees. Similarly, the northern part of Vietnam, referred to as a a non-traditional region for rubber cultivation, where rubber trees have been developed recently, presents various stress factors, particularly climate features such as prolonged low temperature and low solar radiation in the winter, very high air temperature and strong wind in the summer. Until now, the network of clonal trials including 111 trials scattering from 11o N to 22o N latitude are being evaluated. Out of these trials, 60, 16, 11 and 24 trials were located in southeast region, Highlands, Coastal region and Northern region of Vietnam, respectively (Table 4).

Table 4. The network of clonal trials which are being evaluated

Trials Southeast region Highlands Coastal region Northern region

ESTs 5

SCCTs 23 6 5 7

LCCTs and Promotion plots

32 10 6 17

Total 60 16 11 24

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III. 3. Application of biotechnology in Hevea breeding programmes

3.1. Tissue culture

As the conventional method of propagation may lead to undesirable stock-scion interactions, the clonal propagation of Hevea by tissue culture is a relevant technique to a greater extent. The production of uniform individual plants is one of the objectives for in vitro propagation of Hevea. Recently, microcutting and somatic embryogenesis are being carried out in Vietnam. However, the frequency of somatic embryo induction was found to be very low and non-synchronous, its germination remains very difficult and thus Hevea embryogenesis system needs further improvement.

3.2. Genetic markers

Genetic markers have served as useful aids in understanding the genetics of H. brasiliensis in the recent past. In Vietnam, significant progress has been made in application of genetic markers in Hevea breeding programmes. Currently, isozyme electrophoresis, random amplified DNA polymorphism (RAPD) and inter-simple sequence repeat (ISSR) have been used to identify Hevea clones, analyze genetic diversity of Hevea germpasm, and to select good Hevea clones for future breeding or cultivation purposes. The combination of morphological characterization, isozyme, RAPD and ISSR markers could help breeders constitute a core collection of IRRDB’81 germplasm to ensure conserving the genetic variability. In addition, these markers could also help in checking the genetic variability of the Hevea breeding programmes, and could be useful for accumulation and management of genetic-breeding resources of Hevea and would help to utilize the new genetic resources more effectively in Hevea breeding programmes in Vietnam.

3.3. Physiological, biochemical and molecular responses to drought stress

Global warming and climate change involve significant changes in temperature, rainfall and wind. Several studies have highlighted the importance of environmental factors, particularly rainfall which is considered as a major factor that can influence the development of rubber trees by affecting the physiological and biochemical processes in plants. In order to identify the key factors to be used in phenotyping new clones, particularly RRIV’s clones for their tolerance of drought stresses, physiological and biochemical responses and expressions of genes involved in energy biosynthesis and reactive oxygen species (ROS) scavenging, were systematically analysed following drought stress treatments. The results revealed that the combination of physiological, biochemical and molecular parameters are the valuable parameters for early detection of drought stress tolerant clones of Hevea which can be developed at the location of specific clones, especially for drought prone areas in Vietnam.

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The IRRDB Annual Report 2017 – Crop management specialist group

LO : Dr KUSWANHADI

Representatives : CIRAD : Dr Eric GOHET CATAS : Dr An FENG CRRI : Mr Lim Khan Tiva IRRI : Mr JUNAIDI RRIV : Ms Nguyen Thi Hoang Van

RRII : Dr. M.D. Jessy

RRISL : Dr Chamil Nayanakantha RRIM : Ms Arjuna Abang RRIT/RIOT : Mr. Arak Chantuma RRI Philipine : Dr Abello Meynard

INTERNATIONAL RUBBER RESEARCH BOARD 2017

The RRI VIETNAM Issues and challenges

• Continuing decline in the price of natural rubber has resulted in the significantly decreased income of smallholders and low profit of estates.

• Tapping profession is in crisis due to the lack of skilled workers.

• Cost of production is affected by increasing labour wages. Rubber growers are not ready to pay for the high agrochemical input cost.

• Rubber-tree cultivation over a long perios also adversely affects yield.

Strategies and programmes to address issues and challenges

• Advanced latex harvesting technologies such as low intensive tapping system (LITS), low frequency tapping system (LFTS) as well as increased tapping task bring out improved tapper productivity and decreased labour requirement.

• Natural rubber cup lump is offered as an alternative to latex collecting in the areas which have limited tapping labour availability.

A. Latex harvesting systems (research projects)

1. Yield response of several common clones in Viet Nam (PB 260, PB 255, RRIV 114, RRIV 124, RRIV 106) to low frequency tapping systems (d5 and d6). (2017 – 2020)

Objectives: Evaluating the long-term effects of low frequency tapping with increased intensity of stimulation to those clones in different kinds of agro-climatic conditions including the traditional and non-traditional areas. Examining the possible adoption of low frequency tapping systems (d5 and d6) in Viet Nam. Appropriate low tapping frequency is in accord with stimulation regime for each rubber tree clone. Determining the laticiferous physiological status of those clones under different low frequency tapping systems.

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Results: Yield performance of those rubber - tree clones gain under different frequency tapping systems. Adopting low tapping frequencies such as d5 and d6 to rubber estates produces socio-economic effectiveness.

2. Latex collecting management (2017-2020) Objectives : Assessing the natural rubber cup lump collecting in comparison to latex collecting. Results: Field coagula – cup lump can be offered in the non-traditional rubber-growing areas with low numbers of skilled tapping labour.

3. Tapping panel management for upward tapping panel (2015-2018 ) Objectives : Enhancing rubber yield performance over long term under CUT systems (S/4U, S/6U and S/8U) in accordance with stimulant application (ethephon or ethylene stimulation). Results : Appropriate controlled upward tapping systems in relation to the tapping ages of rubber trees.

4. Latex diagnosis (2008-2020 ) Objectives : Determining the under or over latex harvesting status of rubber plantations through the expression of latex physiological parameters. Establishing the latex physiological reference values and reference limits in the latex diagnosis laboratory. Results : Recommendations for appropriate latex harvesting systems in rubber estates and smallholders. Reference intervals for in vitro latex diagnosis under rubber plantations in Southeast region of Viet Nam.

B. Extension 1. Training on latex harvesting technique (LHT) – (2016-2020)

Objectives : Improving the LHT knowledge and tapping skills for tappers as well as plantation managers. Results: 500 - 900 participants annually can be trained.

RRI INDONESIA

1. Low tapping intensity on PB 260 clone Objectives : study the effect of tapping frequency and stimulant application frequency to yield Result : The low tapping intensity increased the yield (g/t/t) significantly compared to control (d3), whilst, among LFT there was no significant yield. The highest yield was found in treatment S/2 d5 ET 2.5% 8/y(m) reached 48% above control 67.11 g/t/t. The LFT tended to have higher DRC although not statistically significant. The increase of stimulant frequency did not decrease the DRC when the tapping frequency reduced. The LFT treatment S/2 d8 ET 2.5% 12/y(m-2w) had a significant lower TPD rate (4.94%). The highest TPD rate was found in control treatment S/2 d3 ET 2.5% 4/y (m) which reached 21.00%. Reducing tapping frequency lower the stress to the plant led to a lower TPD rate (Table 1)

Table 1. Yield, latex DRC, and TPD intensity of various treatment

Tapping frequency Yield (g/t/t) DRC (%) TPD (%)

S/2 d3 ET 2.5%.Ga 4/y (m) 45.35 a 36.67 21.00 b

S/2 d4 ET 2.5%.Ga 6/y (m) 65.41 b 38.33 13.52 ab

S/2 d5 ET 2.5%.Ga 8/y (m) 67.11 b 39.67 5.99 ab

S/2 d6 ET 2.5%.Ga10/y (m) 58.31 ab 40.00 10.78 ab

S/2 d8 ET 2.5%.Ga12/y (2w) 61.86 b 40.67 4.94 a

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2. The Effect of High Stimulant Concentration to Yield and DRC in Low-Tapping Frequency Practice Objectives : study the effect of high concentration of ethepon, PEG and palmitic acid to yield Results : As stimulants were applied once in a month, it would be eight tapping days before they were applied again. The yield pattern exhibited a negative slope to the n-tapping after application. The longer the tapping practice was, the lower yield was obtained. All treatments showed that the first and second tapping after stimulant application generated highest yield, then, it decreased along with the number of tapping. A significant decrease was observed in the third tapping. In ET treatment, the decrease was 30.65% to the second tapping, whilst in EN and NS treatments were 26.12% and 20.40% respectively (Table 2) Table 2. Yield per tapping after stimulant application

n-tapping after stimulant application

Yield (gt-1t-1)

ET EN NS

1 89.61 + 12.32 a 63.38 + 6.01 ab 46.06 + 4.02 a

2 87.83 + 10.49 a 65.23 + 6.36 a 42.59 + 3.57 a

3 60.91 + 8.47 ab 48.19 + 7.55 abc 33.90 + 3.07 b

4 51.87 + 2.22 bc 42.97 + 2.36 bcd 28.58 + 2.01 c

5 51.05 + 3.03 bc 41.78 + 2.83 cd 32.30 + 0.92 bc

6 44.63 + 1.79 cd 35.87 + 2.81 de 32.40 + 1.75 bc

7 42.29 + 2.24 d 33.86 + 2.15 e 29.19 + 1.03 bc

8 39.79 + 2.90 d 33.29 + 3.56 e 32.38 + 1.93 bc

Note : - ET: Etephon 5.0%, - EN: Polyethylene glycol 3.0% + palmitic acid 0.2% + etephon 5.0%, - NS: Polyethylene glycol 3.0% + palmitic acid 2.0%

3. Intercropping in wide spacing double-row rubber

Methods : 5 rubber clones (PB260, PB330, IRR118, IRR220, and BPM24) , 3 spacing, intercropping with banana, sugarcane, palm oil Objectives : increase land productivity and farmers income Results: There is no difference among planting distance (table 3), with a girth of 7.7 cm at 9 Month after planting (MAP). PB260 showed lowest growth rate (Table 4).

Table 3. Plant girth of various planting distance at 9 MAP

Planting distance girth (cm)

20 m x 2 m x 2,3 m 7.8 ns 19 m x 2 m x 2,4 m 7.7 18 m x 2 m x 2,5 m 7.7

Table 4. Plant girth of various hevea clone at 9 MAP

clone girth (cm)

IRR 118 7.7 ab IRR 220 8.0 a PB 260 7.2 b BPM 24 7.8 a PB 330 7.9 a

4. Rubber growing in peat soil

Methods : evaluated 4 planting systems i.e. Control, Deep planting, Deep planting and budding at 30 cm, Deep planting and budding at 50 cm. Objectives : study the possibility of rubber extension to peat-soil

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Results: up to 4 years rubber plant grow well (Figure 1) with a girth of 37 cm for all the treatment (Table.5.)

Figure 1. Plant performance in the field Table 5. Girth of the tree at various treatment

Treatment 48 MAP

Control 37,74 ns Deep planting 37,96 Deep planting and budding at 30 cm 37,76 Deep planting and budding at 50 cm 37,13

RRI INDIA Online fertilizer recommendation (RubSIS) developed by the Rubber Research Institute of India was extended to the entire rubber growing regions in South India. RubSIS was translated to regional languages and is popularized among growers. Follow-up studies of RubSIS was initiated in growers’ fields. To bring the entire rubber growing regions within the ambit of RubSIS, soil sample collection was initiated in the non-traditional rubber growing regions also. A study was conducted to compare the quality of groundwater and surface water in watersheds dominated by different plantation crops viz, rubber, tea and cardamom. Watersheds (of about 25-30 km2 area) dominated by each plantation crop were delineated using the survey of India topographic maps and satellite images. For sampling ground water, about 20-30 dug wells were located which were distributed in the entire watershed area. Similarly, for sampling surface water, sampling points were identified in rivers or major streams. Water quality parameters such as electrical conductivity (EC), salinity, total dissolved solids (TDS), total suspended solids (TSS) and hardness were significantly higher in watersheds of tea and cardamom compared to that in rubber watershed. Contents of potassium, calcium, magnesium, sodium and nitrate showed the same trend. Compared to tea, significantly higher values for these parameters were observed in cardamom watershed. Contents of phosphate, chloride and sulphate were detected in trace quantities. Analysis for heavy metals (lead, cadmium, chromium, zinc and copper) showed that all these were also within permissible limits of drinking water. Dissolved oxygen (DO) content in surface water was significantly higher in the watershed of rubber compared to tea and cardamom, indicating better health of water resource in rubber system. Pesticide residue analysis of representative samples from the three watersheds showed that organo-chlorines, organo-phosphates and pyrithroides were below the level of quantification in ground water and surface water from all three water sheds. Bacteriological analysis showed that total coliforms and e-coli were positive in all the ground water and surface water samples from all the three watersheds.

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CIRAD Experiments followed up by Cirad regarding latex harvest technology are established in partnership with other IRRDB member countries (Universities and research institutes) or with private agro-industry companies. Most of the activities are devoted to the fine optimization of tapping systems recommendations depending on eco-climatic and socio-economic particular contexts and in close connection with rubber physiology and breeding research programmes.

1. Latex Diagnosis (LD)

The most important activity deals with the optimization of tapping systems recommendations using Latex Diagnosis (LD), carried out in two steps.

a. Establishment of basic tapping systems recommendations per any location, depending on clonal typology

b. Local (plot per plot) optimization of these recommendations using Latex Diagnosis (diagnosis or under-exploitation, over-exploitation or normal exploitation, and local corresponding adjustments of tapping systems: increased, decreased or maintained intensity). This relies on an increased capability of LD interpretation permitted by the continuous development of Cirad LD databases and now leads to piloting of rubber plantations tapping intensity on a plot scale (“site specific” recommendations).

This strategy was first presented on the occasion of the IRRDB international workshop on Latex Harvesting Technology, held in Sungai Buloh, Selangor, Malaysia in May 2008 (Gohet et al, 2008) and is consistently being improved, heading towards more and more precise stimulation management. Several Latex Diagnosis laboratories have been installed by Cirad, and this includes staff training for sampling and analyses, inside R&D centres of agro-industrial companies. Experiments in several countries deal with the clonal responses to tapping frequency, stimulation and panel management. As all these experiments are followed up for physiological parameters as well, those help to precise the clonal typology and therefore all tapping systems recommendations. Cirad also continues its huge involvement in the research on optimization of typical smallholders tapping systems, mostly using high tapping frequency resulting from tapping intensification and caused by a continuous decrease of average rubber smallholding size. In such intensive contexts, reduced tapping frequencies and associated compensation by ethephon stimulation are mostly out of reach by planters. General monographies on the Cirad approach to TOT and good agricultural practices have been published during different workshops (Gohet et al 2016 a, b, c, Gohet et al 2017), in Malaysia, Singapore, Thailand and Cambodia. 2. Study of the effect of climate on rubber yield

A study of the effect of climate (temperature and rainfall especially on rubber yield potential has been started by combining our international production database with climatic data, in order to characterize and quantify the intensities of climatic stresses and their effect on rubber yield potential. The ultimate goal is to create a climatic typology of climatic stresses on latex production and to propose related adaptations of optimised tapping systems to reduce these climatic impacts on yields (Gohet et al 2015)

3. Studies on slow release of ethylene gas stimulation

Studies of systems allowing regulation and slow release of ethylene gas stimulation (regulated flow, gas pressure control) are currently under process.

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4. Research on soil quality and sustainability A research programme on soil quality and sustainability has also been started in cooperation with agro-industrial companies in Africa. 5. Recent publications and communications of our team (2016-2018), related to Latex harvesting

Technology and Crop management: Chambon B., Bosc P.M., Promkhambut A., Duangta K. 2018. Entrepreunarial and family business farms in Thaïland: Who took advantage of the rubber boom?. Journal of Asian Rural Study, 2 (2) : p. 173-195. http://pasca.unhas.ac.id/ojs/index.php/jars/article/view/1481 Hougni D.G.J.M., Chambon B., Penot E., Promkhambut A. 2018.The household economics of rubber intercropping during the immature period in Northeast Thailand.Journal of Sustainable Forestry:18 p. https://doi.org/10.1080/10549811.2018.1486716 Kullawong S., Aditto S., Chambon B., Promkhambut A. 2018. Dynamic of rubber production in Northeast Thailand: A case study at Subsomboon village, Doonsard sub-district, Kranuan district, Khon Kaen province. Khon Kaen Agriculture Journal, 46 (1) (suppl.) : p. 239-248. https://ag2.kku.ac.th/kaj/ Penot E., Chambon B., Thériez M., Benavidez López D.N., Tongkaemkaew U. 2018. Les plates formes d'innovation comme nouveau modèle de développement agricole inter-acteurs dans les pays en développement : regard croisé sur deux expériences en cours en Thaïlande et au Nicaragua. In : Les nouveaux modes d'organisation des processus d'innovation. Montpellier : RRI, 20 p. Congrès RRI - Forum innovation. 8, 2018-06-04/2018-06-05, Nîmes (France). Stroesser L., Penot E., Michel I., Tongkaemkaew U., Chambon B. 2018. Income diversification for rubber farmers through agroforestry practices. How to withstand rubber price volatility in Phatthalung Province, Thailand. Revue Internationale des Etudes du Développement (235) : 24 p. http://www.publications-sorbonne.fr/fr/revues/ried/ Tongkaemkaew U., Chambon B. 2018. Rubber plantation labor and labor movements as rubber prices decrease in southern Thailand. Forest and Society, 2 (1) : p. 18-27. https://doi.org/10.24259/fs.v2i1.3641 Chambon B., Lai Dao X., Tongkaemkaew U., Gay F. 2017. What determine smallholders' fertilization practices during the mature period of rubber plantations in Thailand?. Experimental Agriculture : 18 p. http://dx.doi.org/10.1017/S0014479717000400 Chambon B., Promkhambut A., Duangta K., Lesturgez G., Sainte-Beuve J. 2017. Practices and quality perceptions of farmers producing cup coagulum in Thailand. Rubber Science, 30 (3) : p. 213-225. Chantuma P., Lacote R., Sonnarth S., Gohet E. 2017. Effects of Different Tapping Rest Periods during Wintering and Summer Months on Dry Rubber Yield of Hevea Brasiliensis in Thailand. Journal of Rubber Research, 20 (4) : p. 261-272. http://rios.lgm.gov.my/cms/fedDigiJournalDetail.jsp?searchText=&selTab=digiCon&id=&type=JRR&issueYear=0 Gohet E., Lacote R., Leconte A., Chapuset T., Rivano F., Chambon B. 2017. Improving Rubber Smallholdings Productivity and Resilience through Adoption of Good Agricultural Practices. In : CRRI-IRRDB Workshop on Transfer of Technology to Rubber Growers ¿Good Agricultural Practices Reduce Wastages, Improve Quality, Productivity and Profitability¿. Phnom Penh : CRRI-IRRDB, p. 1-15. CRRI-IRRDB Workshop on Transfer of Technology to Rubber Growers ¿Good Agricultural Practices Reduce Wastages, Improve Quality, Productivity and Profitability¿, 2017-07-12/2017-07-14, Phnom Penh (Cambodge). Lerksamran T., Clément-Vidal A., Liengprayoon S., Rattanaporn K., Gohet E., Thaler P., Chantuma P., Lacote R. 2017. Relationships between carbon stock in the rubber tree and latex production. Jakarta : IRRDB, 9 p. International Rubber Conference & IRRDB Annual Meeting 2017, 2017-10-18/2017-10-20, Jakarta (Indonésie). Nicod T. 2017. Etude sur les stratégies d'adaptation des petits planteurs d'hévéa Thaïlandais, face à la chute du prix du caoutchouc naturel. Cergy-Pontoise : ISTOM, 99 p. Mémoire de fin d'études -- , Mémoire de fin d'études -- , Mémoire de fin d'études -- . Penot E., Chambon B., Wibawa G. 2017. History of rubber agroforestry systems development in Indonesia and Thailand as alternatives for sustainable agriculture and income stability. s.l., 26 p. IRRDB 2017, 2017-10/2017-10, Bali (Indonésie). Penot E., Snoeck D., Vagneron I. 2017. Creating shared values in agroforestry systems based on coffee and cocoa clusters. Exploratory mission: mission report. Montpellier : CIRAD, 64 p. Sainoi T., Sdoodee S., Lacote R., Gohet E. 2017. Low frequency tapping systems applied to young-tapped trees of Hevea brasiliensis (Willd. ex A. Juss.) Müll. Arg. in Southern Thailand. Agriculture and Natural Resources, 51

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(4) : p. 268-272. http://dx.doi.org/10.1016/J.ANRES.2017.03.001 Sainoi T., Sdoodee S., Lacote R., Gohet E., Chantuma P. 2017. Stimulation affecting latex physiology and yield under low frequency tapping of rubber (Hevea brasiliensis) clone RRIM 600 in southern Thailand. Australian Journal of Crop Science, 11 (2) : p. 220-227. http://dx.doi.org/10.21475/ajcs.17.11.02.p305 Samila Y., Ayutthaya S.I.N., Meetha S., Chantuma P., Lacote R. 2017. Use of ethylene stimulation to enhancing the rubber tapping in northeast Thailand. Khon Kaen Agriculture Journal, 45 (1) (suppl.) : p. 321-324. https://ag2.kku.ac.th/kaj/ Thaler P., Desalme D., Duangngam O., Kasemsap P., Sathornkich J., Chayawat C., Satakhun D., Priault P., Angeli N., Chantuma P., Epron D. 2017. Tracking carbon from photosynthesis to latex with 13C field labelling experiment. s.l., 1 p. IRRI and IRRDB International Rubber Conference, 2017-10-18/2017-10-20, Jakarta (Indonésie). Thériez M., Penot E., Tongkaemkaew U., Chambon B. 2017. De l'indifférence à la reconnaissance : potentiel de développement d'une plateforme d'innovation régionale sur les systèmes agroforestiers hévéicoles du sud de la Thaïlande. Paris : SFER, 28 p. Journées de Recherches en Sciences Sociales SFER. 11, 2017-12-14/2017-12-15, Lyon (France). https://www.sfer.asso.fr/event/view/10 Thoumazeau A., Gay F., Alonso P., Suvannang N., Panklang P., Chevallier T., Bessou C., Brauman A. 2017. SituResp®: A time- and cost-effective method to assess basal soil respiration in the field. Applied Soil Ecology, 121 : p. 223-230. http://dx.doi.org/10.1016/j.apsoil.2017.10.006 Vaysse L., Sathornluck S., Bottier C., Liengprayoon S., Char C., Bonfils F., Chotiphan R., Lacote R., Gay F. 2017. Effect of fertilization and stimulation of Hevea brasiliensis trees on mineral compositions and properties of produced latex and rubber. Jakarta : IRRDB, 9 p. International Rubber Conference & IRRDB Annual Meeting 2017, 2017-10-18/2017-10-20, Jakarta (Indonésie). Zar Ni Zaw, Sdoodee S., Lacote R. 2017. Performances of low frequency rubber tapping system with rainguard in high rainfall area in Myanmar. Australian Journal of Crop Science, 11 (11) : p. 1451-1456. http://dx.doi.org/10.21475/ajcs.17.11.11.pne593 Chambon B., Promkhambut A., Tongkaemkaew U., Lai Dao X., Bosc P.M. 2016. Has the rubber boom weakened households in the new rubber producing areas? A case study in Northeast Thailand. In : CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 1-13. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Chambon B., Ruf F., Kongmanee C., Angthong S. 2016. Can the cocoa cycle model explain the continuous growth of the rubber (Hevea brasiliensis) sector for more than a century in Thailand?. Journal of Rural Studies, 44 : p. 187-197. http://dx.doi.org/10.1016/j.jrurstud.2016.02.003 Chantuma P., Somnarth S., Lacote R., Gohet E. 2016. Result of the study DCA (double cut alternative tapping system) and Single cut with RRIM 600 over a period of 15 tapping years. Para Rubber Bulletin, 37 (1) : p. 4-12. Chapuset T., Anleu Morales J.V., Snoeck D., Palu S., Najera C.A. 2016. Improving rubber tree productivity by reducing tapping frequencies in Guatemala. In : Berti M.T. (ed.), Alexopoulou E. (ed.). Industrial crops: promoting sustainability. Rochester : AAIC, p. 75-75. Association for the Advancement of Industrial Crops International Conference, 2016-09-24/2016-09-28, Rochester (Etats-Unis). http://www.aaic.org/past-meeting-program-and-abstracts.html Gohet E., Chambon B., Lacote R. 2016. Optimizing smallholders' yield and productivity through adoption of appropriate latex harvesting technology. In : MRB IRRDB Workshop on Agronomy and Transfer of Technology. Sungai Petani : IRRDB, 15 p. MRB IRRDB Workshop on Agronomy and Transfer of Technology, 2016-04-26/2016-04-27, Sungai Petani (Malaisie). http://www.lgm.gov.my/news/news2016AprMRB-IRRDBWorkshop.htm Gohet E., Lacote R., Leconte A., Chapuset T., Rivano F., Chambon B. 2016. Improving rubber smallholdings productivity and resilience through adoption of good agricultural practices. Singapour : s.n., 15 p. Focus Forum on Natural Rubber Sustainability, 2016-05-10/2016-05-11, Singapour (Singapour). Gohet E., Lacote R., Leconte A., Chapuset T., Rivano F., Chambon B. 2016. Optimizing smallholders yield through adoption of good agricultural practices. In : Global Rubber Conference 2016 : Driving Transformation and Unlocking Opportunitie¿. Krabi : IRRDB, p. 1-15. Global Rubber Conference 2016, 2016-10-11/2016-10-13, Krabi (Thaïlande).

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Lim Khan Tiva, Lacote R., Chhek C., Mak S., Gohet E. 2016. Effect of fertilization on latex yield potential and physiological parameters of clone PB 217 in Cambodia. In : CRRI, IRRDB. CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 409-414. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Penot E., Stroesser L., Michel I., Tongkaemkaew U., Chambon B. 2016. Agroforestry practices to overcome prices volatility: the case of rubber in Phatthalung province, Thailand. Abstract number 92. In : Gosme Marie (ed.). Celebrating 20 years of agroforestry research in Europe. Book of Abstracts. Montpellier : EURAF, p. 169-172. European Agroforestry Conference. 3, 2016-05-23/2016-05-25, Montpellier (France). http://www.agroforestry.eu/conferences/III_EURAFConference

Phean Chetha, Phearun P., Hak Bunthuon, Lacote R., Chhek C., Mak S., Gohet E. 2016. Yield performance of 8 rubber clones after 14 years of tapping in Cambodia. In : CRRI, IRRDB. CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 281-285. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Phearun P., Chetha P., Bunthuon H., Lacote R., Chhek C., Mak S., Gohet E. 2016. Early clonal selection of Hevea brasiliensis based on latex physiological parameters in Cambodia. In : CRRI, IRRDB. CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 286-297. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Rivano F., Gohet E., Lacote R., Leconte A., Chapuset T., Chambon B. 2016. Mejorar la productividad de las plantaciones de caucho implementando buenas prácticas agrícolas. Bucaramanga : Confederación Cauchera Colombiana, 1 p. Congreso Internacional Cauchero - Eficiencia y Productividad, un Camino Hacia la Paz. 6, 2016-10-13/2016-10-15, Bucaramanga (Colombie). Rivano F., Vera J., Cevallos V., Almeida D., Maldonado L., Flori A. 2016. Performance of 10 Hevea brasiliensis clones in Ecuador, under South American Leaf Blight escape conditions. Industrial Crops and Products, 94 : p. 762-773. http://dx.doi.org/10.1016/j.indcrop.2016.09.035 Sainoi T., Sdoodee S., Lacote R., Gohet E., Chantuma P. 2016. Assessment of low frequency tapping systems with stimulation on Hevea in Southern Thailand. In : CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 172-182. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Sengkea H., Gohet E., Chhek C., Mak S., Lacote R. 2016. Yield potential of clone IRCA 230 in Cambodia. In : CRRI, IRRDB. CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 269-280. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). Stroesser L., Penot E., Michel I., Tongkaemkaew U., Chambon B. 2016. Income diversification for rubber farmers through agro-forestry practices: how to overcome rubber prices volatility in Phatthalung province, Thailand. In : CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 76-97. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html Thaler P., Chambon B., Bosc P.M., Penot E., Robain H., Do F.C., Gay F., Tongkaemkaew U., Suvannang N., Trisophon K., Sajjaphan K., Kasemsap P. 2016. Heveadapt: How can Thai rubber smallholders face global challenges?. Xishuangbanna : s.n., 1 p. Sustainable Rubber Conference 2016, 2016-10-16/2016-10-19, Xishuangbanna (Chine). http://sustainablerubber.org/conference-2016/abstracts/#15 Thoumazeau A., Bessou C., Gay F., Thaler P., Trap J., Phongjinda A., Suvannang N., Sajjaphan K., Brauman A. 2016. "Biofunctool": a framework to assess the impact of agricultural practices on soil quality based on soil functions. Montpellier : INRA, IRD, 2 p. International EcoSummit Congress 2016, 2016-08-29/2016-09-01, Montpellier (France). Tongkaemkaew U., Chambon B. 2016. Labour used in rubber plantations and labour movement in rural, sub-urban and urban communities: a case study in Songkhla Province, Southern Thailand. In : CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 50-60. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html

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Zar Ni Zaw, Sdoodee S., Lacote R. 2016. Preliminary study on low intensity tapping system with rainguard in a high rainfall area in Myanmar. In : CRRI, IRRDB. CRRI and IRRDB International Rubber Conference 2016, Siem Reap, Cambodia. Siem Reap : CRRI, IRRDB, p. 230-240. International Rubber Conference, 2016-11-21/2016-11-25, Siem Reap (Cambodge). http://rri.catas.cn/en/news/?106.html

CATAS 1. Low frequency and low intensity tapping system

Low frequency and low intensity tapping system of 4d and 5d +ET tapping system were widely implemented. Meanwhile, 1/8 S+ Ethylene gas and 7d +ET tapping system were extended in some farms and Acupunture+ Ethylene gas stimulated tapping system was examined. 2. Electronic tapping knife

A newly developed electronic tapping knife (4GXJ-I) was in mass production and extended to some rubber plantations in China, Myanmar, Laos, Vietnam, Thailand, Cambodia, Sri Lanka, Malaysia and Indonesia (Fig 2). Fig.2. Electronic tapping knife (4GXJ-I)

3. Planting Material

In the integrated tube training technique for producing budded rubber saplings, the bio-degradable tubes were used. Thus, the grafted rubber tree saplings can be planted without removing the tube and the tube can be degrade in less than six months

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Figure 3 Meanwhile, a maximum-stump rubber sapling producing technique was developed to reduce gestation period of the rubber tree. The sampling grows very well six months after planting (Figure 4)

Figure 4 4. TPD control

TPD control TPD recovery nutrient agent was registered by the Chinese government and was produced in mass production and implemented in large-scale.(Figure 5)

Figure 5

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5. Fertilizer Formulation

To save the labor some new cover materials and fertilizer bars were utilized. (Figure 6) Fig 5

Figure 6 6. Rubber based cropping system

To make the rubber plantations more productivity and profitable, the 2×4+20m rubber based cropping system was more widely extended (Fig 7). More crop, vegetable, cash crop and patterns were tested.

Figure 7

Figure 7a.

Figure 7b

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RRI SRI LANKA Research activities conducted at Rubber Research Institute of Sri Lanka and other universities of Sri Lanka with regard to planting material production, field establishment, immature upkeep tapping, tapping panel dryness and intercropping are reported here. A. Rubber Research Institute of Sri Lanka, Agalawatta, Sri Lanka 1. Nursery inspection Government, Regional Plantaion Companies and Private nurseries were inspected for quality assurance (N M C Nayanakantha, B M S S Panditharathna, M N de Alwis , L N de Zoysa and R Handapangoda) 2. An assessment on the vulnerability of Hevea seed production to climate change (CC/2003/1) Wintering and flowering (CC/2003/1/a) Wintering and flowering were observed for 26 clones at five estates and one small holder field during January-February. Majority of the clones completed wintering during 2nd-3rd week of February. (N M C Nayanakantha and P D Pathirana) 3. Seed production (CC/2003/1/b) Seeds of 26 clones were collected from six estates and one small holder field (Figure 8). A slight increase in seed production was recorded for 2017 than for 2016 irrespective of clones and sites.

Fig. 8. Effect of clone and site on seed production of rubber 5. Investigation on alternative and cost-effective sowing media for river sand for germination of

rubber seeds (SM/ 2016/Dartonfield)

Rubber seeds brought to the government rubber nursery, Gurugoda, were sown in different sowing media as depicted below; Control : River sand Treatment 1 : Elephant dung (dried) Treatment 2 : Coir dust (leached) Treatment 3 : Rubber wood chips Treatment 4 : River sand + coir dust (leached) (1:1)

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Treatment 5 : Rubber wood saw dust Treatment 6 : River sand + Purified sea sand (1:1) Treatment 7 : Gravel Treatment 8 : Quarry dust (fine) Treatment 9 : Purified sea sand After three weeks of sowing, a satisfactory germination was recorded from seeds sown in elephant dung or coir dust as compared to river sand (Figure 9.).

Fig. 9. Effect of sowing media on germination of rubber seeds after 21 days of sowing. 6. Priming of rubber seeds for improved germination dynamics, seedling and budded plant

attributes and tolerance to abiotic stresses (ST/2013/DF)

Fresh rubber seeds of clone RRISL 217 were collected and soaked in nitric oxide (NO) donor sodium nitroprusside (SNP) solutions at different concentrations for 24 hours as shown below; T1 = Control (no soaking) T2 = Soaked in water T3 = Soaked in 50µmol SNP T4 = Soaked in 100µmol SNP T5 = Soaked in 150µmol SNP

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Fig 10. Effect of NO donor SNP on germination percentage of rubber seeds after 0 day (A), 7 days (B), 14 days (C) and 21 days (D) of storage. Means with same letter are not significantly different at p≤0.05 according to DMRT. After the 7thday of storage, a significantly (p≤0.05) higher germination percentage was recorded with SNP at 50 µM [80. 7% (after 7 days) and 99.2% (after 14 days) of sowing ] as compared to the control [ (60% (after 7 days) and 75.7% (after 14 days) of sowing]. After the 14th and 21st day of storage, a significantly (p≤0.05) higher germination percentage was recorded with SNP at 50 and 100 µM when compared to the control and mock treatment after 21 days of sowing (Figure 10.). Table 6. Effect of exogenous NO donor SNP on girth of Hevea after three months from treatment imposition. 7. Irrigation systems for rubber nurseries

Effect of exogenous application of salicylic acid on physiological and morphological attributes of nursery-gown rubber plants under water stress A polybag nursery was established at Moneragala substation to optimize the application interval of salicylic acid at a concentration of 0.5 mM with different irrigation systems. W/o SA – without application of salicylic acid SA 2 weeks – Application of salicylic acid once in two weeks SA 1 month– Application of salicylic acid once a month SA 3 months – Application of salicylic acid once in three months applications

Treatment Girth (cm) at Day 0 Girth (cm) after 3 months

Control 4.54±0.11 5.78±0.08 Water 4.44±0.07 6.43±0.48 100 µM SNP 4.27±0.10 6.20±0.14 150 µM SNP 4.54±0.19 6.25±0.10 200 µM SNP 4.45±0.06 6.53±0.31* LSD0.5 0.3205 0.7187

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Table 7. Growth, physiological and bud grafting performance of plants after five months of treatment imposition

Treatments Stem diameter (mm)

Leaf chlorophyll content (SPAD unit)

Stomatal conductance (mm/s)

Bud grafting success (%)

Sprinkler w/o SA 7.85± 0.13b 56.88 ±1.70 a 0.11± 0.02a 84.2 ± 0.82b

SA 2 weeks 7.92 ± 0.10 b 56.06 ± 1.36a 0.10 ± 0.03a 84.6 ± 0.90b

SA 1 month 8.35 ±0.08a 56.26 ±1.85a 0.13 ± 0.02a 91.6 ± 0.45a

SA 3 months 8.00 ± 0.11b 56.44 ±1.43a 0.11± 0.04a 90.0 ± 0.85a

Drip w/o SA 6.80 ± 0.10bc 52.86 ± 1.29ab 0.05 ± 0.02bc 79.0 ± 0.74c

SA 2 weeks 7.40 ± 0.10bc 53.08 ± 1.07ab 0.01 ± 0.02c 80.2 ± 0.56bc

SA 1 month 7.87 ± 0.11b 54.56 ±1.62a 0.03 ± 0.03c 83.3 ± 0.52b

SA 3 months 7.53 ± 0.09bc 55.48 ± 1.42a 0.02 ± 0.03c 81.5 ± 0.72b

Manual w/o SA 6.46 ± 0.10c 51.54 ± 1.04b 0.05 ± 0.02bc 82.5 ± 0.90b

SA 2 weeks 6.55 ± 0.08bc 51.60± 0.67b 0.09 ± 0.01b 80.4 ± 0.72bc

SA 1 month 7.36 ± 0.11b 52.48 ± 0.64ab 0.13 ± 0.02a 85.2 ± 0.65b

SA 3 months 7.08 ± 0.08bc 51.84 ± 0.60b 0.11 ± 0.02a 85.0 ± 0.59b

-Means with the same letter are not significantly different at p <0.05 8. Northern Province Planting (PT/2017/Kilinochchi)

New experimental fields were established at Malayalapuram, Kilinochchi, Dry Zone, in four farmer fields. Extent of each field was about 0.5 to 1 Acre. Different spacing systems and clones were tested under non - irrigated and irrigated conditions. RRIC 100, RRIC 121, RRIC 102 and RRISL 203 were planted at different spacings of 10’x14’, 8’x14’, 10’x 15’, 13’x13’ and 8’x26’ (control)(N M CNayanakantha, S A Nakandala, B M S SPanditharathne, M N de Alwis and D L N de Zoysa). 9. Stumped buddings experiment (SB/ 2016/Moneragala)

An experimental field was established with RRIC 121 in November 2016 at Moneragala substation and maintenance, agromanagement practices and recording of girth were done. (N M C Nayanakantha, S ANakandalaand L N de Zoysa). 10. Latex Harvesting System

a. Longer tapping cycle through shorter tapping cuts – Pititakanda Estate

Yield measurements for the period from January to December 2017 are given in Table 8.

Table 8. Yield measurements for the period from January to December 2017

Treatments No. of trees

No. of tapping days

Average g/t/t

BB %

T1 : S/4 d3 + 5% E 12/y 42 10 14.6a 4 T2 : S/3 d3 + 2.5% E 12/y 44 10 14.0a - T3 : S/2 d3 + 2.5% E 5/y (* BUT up to December 2010)

43 10 13.9a 4

T4 : S/2 d3 + 2.5% E 5/y 39 10 14.9a 4

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Same yield could be obtained by consuming less bark in T1 and T2 as compared to control (T4)(N M C Nayanakantha, K S D N Senanayake, R K Samarasekera and C Jayawanthi). b. Crop estimation experiment – Sapumalkanda Estate (CE/2012/Sapumalkanda)

Girth, bark thickness, bark consumption and yield measurements were recorded and summaries are given in Table 9. Table 9. Mean girth, bark thickness, annual bark consumption and yield measurements for the period from January to December 2017

Girth class (cm)

Girth (cm)

Bark thickness (mm)

Annual bark consumption (cm)

g/t/t

(51-55.9) 63.0 8.1 a 14.0 67.1 a (56-60.9) 68.8 8.6 a 13.9 53.0 a

≥(61.0) 72.6 8.5a 13.1 65.2 a

There was no significant difference in g/t/t among the girth classes tested (N M C Nayanakantha, P Seneviratne and R K Samarasekara) c. Tapping Panel Dryness

Testing remedies to address tapping panel dryness problem (TPD/2016)

Application of chemicals and botanicals having antioxidant properties on rubber trees Objective of this experiment was to study the effect of bioregulators [chemicals or botanicals (natural plant extracts)] having antioxidant properties on alleviating TPD incidence. Both healthy and TPD affected trees were selected and application of different bioregulators were done on tapping panels. d. Impact of different bark consumption rates associated with additional days of latex harvesting

on growth, yield and economical implications of rubber plantations

This study was planned to determine the impact of different bark consumption rates associated with additional days of latex harvesting (over the recommended) on growth, yield and financial implications of rubber plantation. e. Novel approach to optimize the bark management of rubber plantations (BCR/2017/Sirikandura)

Considering the challenges faced by both smallholder and plantation sector at present, this study was planned to address some issues such as daily tapping, high wage rates to be paid for tappers and imposition of high stress on middle aged rubber trees for obtaining more crop. Therefore, the objective of this study was to find ways and means to increase the productivity of rubber plantations by minimizing bark consumption rates during tapping. A rubber field (2 ha of RRIC 121) established in 2005 at Sirikandura estate, Kalutarawas selected. Initial growth and yield measurements were taken before imposing treatments as shown below;

Treatment Tapping system

T1 T2 T3 T4 T5 T6

S/2 d2 + Recommended No. of Recovery Tapping (Control) S/2 d1 + without rainguard, no recovery tapping (Smallholder Practice) S/4 d1 + With RG (No RT) S/2 d2 (RG) No RT + Supplementary Holiday tapping (S/4 U d7) S/4U d3 2.5% ET + S/2 D d3 2.5%ET (12M alternatively) S/2 D d6 5%ET (Monthly)

(T U K Silva , P Seneviratne, and H Subasingha)

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11. Intercropping Agarwood with Rubber (IC/AW/2015)

Girth data was recorded at two months intervals from January to December 2017 (Table 10). The highest girth and girth increment were recorded from A. crassna and the lowest was recorded from G. walla in both double and single row systems under full sun light condition. Under the natural shading system, the highest girth was recorded from A. crassna and girth increment was as same in A. crassna and A. subintegra. Table 10. Growth attributes of three agarwood species after 28 months from planting. Gyrinops walla (GW), Aquilaria crassna (AC) and Aquilaria subintegra (AS) grown as intercrops with rubber under three planting system

Planting system of Rubber

Agarwood Species Girth at Planting (cm) Girth increment(cm)

Double Row

AC 29.90 ± 0.62 2.59±0.32

GW 9.64±0.51 0.94±0.08

AS 26.70±0.88 1.93±0.30

Single Row

AC 27.61± 1.26 2.44±0.37

GW 7.16±0.66 0.86±0.12

AS 26.53±1.16 2.54±0.32

Under natural shade

AC 14.74± 0.99 0.83±0.31

GW 7.06±0.91 0.52±0.10

AS 13.08±1.36 0.83±0.34

B. University of Peradeniya, Peradeniya, Sri lanka

• Early detection and identification of the causal factors for Tapping Panel Dryness

C. Wayamba University of Sri Lanka, Makandura, Sri Lanka

• Growth and Stand/ ha variations in relation to Clonal, climatic and management effects.

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International Rubber Research and Development Board Molecular Biology and Physiology Specialist Group

Annual Report 2017 Keng-See Chow, Han Cheng, Sigit Ismawanto, Dian Kouadio, Julie Leclercq, Minami Matsui, Thitaporn Phumichai, Thakurdas Saha, Roslinda Sajari, Dinh Minh Tran HIGHLIGHTS IRRDB biotechnology activities in the year 2017 revealed continuing focus on rubber genome sequencing. Following the publication of two high quality rubber genome drafts by CATAS, China (Tang et al., Nature Plants 2016) and RIKEN, Japan/USM, Malaysia (Lau et al., Scientific Reports 2016), a third genome assembly was published by Thailand in 2017 (Pootakham et al., Scientific Reports 2017). Together with other unpublished rubber genome initiatives, six rubber pedigree clones have now been sequenced to varying degrees: Reyan-7-33-97 (China), RRIM 600 (Japan/ Malaysia), BPM 24 (Thailand), RRIM 928 (MRB, Malaysia), RRII 105 (RRII, India) and PB 260 (Bridgestone Co., Japan). With the availability of genome resources and genomic technology platforms, rubber is fast entering the post-genome era of biotechnology. This is reflected by the nature and trend of approaches undertaken by various IRRDB members. As shown in this Annual Report, an increasing number of journal papers is reporting genome-guided discovery and characterization of gene families with functions impacting tree physiology and productivity. Highly saturated genetic linkage maps may be anticipated from more than one group in the near future for application in trait gene/QTL mapping and marker-assisted breeding. Utilization of SNP and SSR markers is noted for gene-specific inheritance studies, clone genotyping and diversity analysis of genetic resources. In addition, attention on phenotyping traits such as Tapping Panel Dryness and molecular weight distribution shows the importance of accurate trait quantification for studying genetic regulation, and inheritance patterns in appropriate mapping populations. Member countries continue to share overlapping goals in many physiological investigations, notably Tapping Panel Dryness, responses in latex to exogenous ethylene, abiotic stress and disease resistance. As has long been noted, formulation of joint projects of common interests under the banner of the IRRDB, including mutual agreement on IP sharing, are much needed to promote efficient collaborations. A Rubber Genomics and Molecular Genetics Workshop was held at CATAS, China from 27-29 September 2017. The Workshop attracted participants from China, India, Indonesia, Japan, France, Malaysia and Thailand. A notable achievement on implementation of the first GWAS analysis of latex yield and girth in an Amazonian population was presented by Thailand. RIKEN, Japan reported the construction of a new Rubber Transcriptome DB. A roundtable discussion at the end of the Workshop highlighted the importance of constructing a unified Hevea Database for rubber molecular resources and the need to fast-track tools for molecular breeding. Subsequently, implementation of a Hevea Database project by CATAS, China was approved at the IRRDB Board of Directors meeting in October, 2017. The Molecular Biology and Physiology Specialist group underwent administrative and membership changes in 2017. Dr. Keng-See Chow was appointed as the Liaison Officer by the IRRDB Board of Directors meeting in October, 2017. RIKEN, Japan became an official member of the IRRDB and the Specialist Group welcomed Prof. Minami Matsui, representing Japan. With that, this Specialist Group takes pride in having members from the three rubber genome publications (2016-2017), and thus looks forward to closer scientific co-operation.

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CHINA Sequencing of the hardy rubber tree genome, Eucommia ulmoides Wuyun Ta-Na, Gao Ruiwen, Li Dejun, Du Hongyan (Chinese Academy of Forestry, RRI-CATAS, BELO EUCOMMIA) A high-quality assembly of its ∼1.2 Gb genome (scaffold N50 = 1.88 Mb) with at least 26,723 predicted genes for E. ulmoides, the first sequenced genome of the order Garryales. As a sister taxon to lamiids and campanulids, the E. ulmoides underwent an ancient genome triplication shared by core eudicots but no further whole-genome duplication (WGD) in the last ∼125 million years. E. ulmoides exhibits high expression levels and/or gene number expansion for multiple genes involved in stress responses and biosynthesis of secondary metabolites, which may account for environmental adaptability. In contrast to rubber tree (Hevea brasiliensis), which produces cis-polyisoprene, E. ulmoides has evolved to synthesize long-chain trans-polyisoprene via farnesyl diphosphate synthases (FPSs). Moreover, FPS and rubber elongation factor/small rubber particle protein gene families were expanded independently from the H. brasiliensis lineage. Transcript profiling of Hevea brasiliensis during latex flow Chao Jinquan, Yang Shuguang, Chen Yueyi, Tian Weimin (RRI-CATAS). The expression levels of 38 genes were significantly higher in CATAS8-79 latex than in CATAS7-33-97 during latex regeneration, while 45 genes had a notably higher expression level in CATAS8-79 latex during latex flow. Together with the activation of the MEP pathway and jasmonate pathway in CATAS8-79 latex, HbPIP1;3, HbPIP1;4, HbSUT3, HbSus3, HbHMGS1-2, HbMK should contribute to the high latex regeneration ability. The up-regulation of ethylene signaling and Hb44KD and the down-regulation of latex coagulation-related genes in CATAS8-79 latex might contribute to its longer latex flow duration. The SWEET gene family in Hevea brasiliensis Sui Jin-Lei, Xiao Xiao-Hu, Qi Ji-Yan, Fang Yong-Jun, Tang Chao-Rong (RRI-CATAS, Hainan University) There are 36 members of the SWEET gene family in this species, making it one of the largest families in plant genomes sequenced so far. Structure and phylogeny analyses of these genes in Hevea and in other species demonstrated broad evolutionary conservation. RNA-seq analyses revealed that SWEET2, 16, and 17 might represent the main evolutionary direction of SWEET genes in plants. Our results in Hevea suggested the involvement of HbSWEET1a, 2e, 2f, and 3b in phloem loading, HbSWEET10a and 16b in laticifer sugar transport, and HbSWEET9a in nectary-specific sugar transport. Parallel studies of RNA-seq analyses extended to three other plant species (Manihot esculenta, Populus trichocarpa, and Arabidopsis thaliana) produced findings which implicated MeSWEET10a, 3a, and 15b in M. esculenta storage root development, and the involvement of PtSWEET16b and PtSWEET16d in P. trichocarpa xylem development. RT-qPCR results further revealed that HbSWEET10a, 16b, and 1a play important roles in phloem sugar transport. Cysteine protease-encoding genes homologous in Hevea genome Zou Zhi, Liu Jianting, Yang Lifu, Xie Guishui (RRI-CATAS) This study presents the identification of SAG12-like genes from the rubber tree genome. Results showed that an unexpected high number of 17 rubber orthologues with a single intron were found, contrasting the single copy with two introns in Arabidopsis. The gene expansion was also observed in another two Euphorbiaceae plants, castor bean (Ricinus communis) and physic nut (Jatropha curcas), both of which contain 8 orthologues. In accordance with no occurrence of recent whole-genome duplication (WGD) events, most duplicates in castor and physic nut were resulted from tandem duplications. In contrast, the duplicated HbSAG12H genes were derived from tandem duplications as well as the recent WGD. Expression analysis showed that most HbSAG12H genes were lowly expressed in examined tissues except for root and male flower. Furthermore, HbSAG12H1 exhibits a strictly senescence-associated expression pattern in rubber tree leaves, and thus can be used as a marker gene for the study of senescence mechanism in Hevea.

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S-adenosylmethionine decarboxylase gene in rubber tree Zhao Manman, Liu Hui, Deng Zhi, Chen Jiangshu, Yang Hong, Li Huiping, Xia Zhihui, Li Dejun (RRI-CATAS, Hainan University, Huazhong Agricultural University) An SAMDC gene (HbSAMDC1) was characterized in rubber tree. HbSAMDC1 contains a 1,080 bp open reading frame (ORF) encoding 359 amino acids. Quantitative real-time PCR analyses revealed that HbSAMDC1 exhibited distinct expression patterns in different tissues and was regulated by various stresses, including drought, cold, salt, wounding, and H2O2 treatments. HbSAMDC1 5' untranslated region (UTR) contains a highly conserved overlapping tiny and small upstream ORFs (uORFs), encoding 2 and 52 amino acid residues, respectively. No introns were located in the main ORF of HbSAMDC1, whereas two introns were found in the 5' UTR. In transgenic tobaccos, the highly conserved small uORF of HbSAMDC1 is found to be responsible for translational repression of downstream β-glucuronidase reporter. Papain-like cysteine protease encoding genes in rubber tree Zou Zhi, Xie Guishui, Yang Lifu (RRI-CATAS) This study performed a genome-wide identification of PLCP family genes in rubber, resulting in a relatively high number of 43 members. The phylogenetic analysis assigned these genes into nine subfamilies, i.e. RD21 (6), CEP (4), XCP (4), XBCP3 (2), THI (1), SAG12 (18), RD19 (4), ALP (2), and CTB (2). Most of them were shown to have orthologs in Arabidopsis; however, several members in SAG12, CEP and XBCP3 subfamilies form new groups as observed in other core eudicots such as Manihot esculenta, Ricinus communis, Populus trichocarpa, and Vitis vinifera. Based on an expert sequence comparison, 20 orthologous groups (OGs) were proposed for core eudicots, and rubber paralogs were shown to be resulted from the recent whole-genome duplication (WGD) as well as local duplication. Transcriptional profiling showed distinct expression pattern of different members across various tissues, e.g. root, leaf, bark, laticifer, flower and seed. By using the senescence-specific HbSAG12H1 as the indicator, the transcriptome of senescent rubber leaves was deeply sequenced and several senescence-associated PLCP genes were identified. Identification and characterization of the abscisic acid (ABA) receptor gene family Guo Dong, Zhou Ying, Li Hui-Liang, Zhu Jia-Hong, Wang Ying, Chen Xiong-Ting, Peng Shi-Qing.(ITBB-CATAS) Fourteen ABA receptor PYL proteins/genes (designated HbPYL1 through HbPYL14) were identified in the most recent rubber tree genome. A phylogenetic tree was constructed, which demonstrated that HbPYLs can be divided into three subfamilies that correlate well with the corresponding Arabidopsis subfamilies. Eight HbPYLs are highly expressed in laticifers. Five of the eight genes are simultaneously regulated by ABA, jasmonic acid (JA) and ethylene (ET). Multiple protein species of REF/SRPP family respond diversely to ethylene stimulation Tong Zheng, Wang Dan, Sun Yong, Yang Qian , Meng Xueru, Wang Limin, Feng Weiqiang, Li Ling, Wurtele Eve Syrkin, Wang Xuchu (ITBB-CATAS, Hainan University, Mississippi State University, Iowa State University) Sequence analysis showed that REFs have a variable and long N-terminal, whereas SRPPs have a variable and long C-terminal beyond the REF domain, and REF258 has a β subunit of ATPase in its N-terminal. Through two-dimensional electrophoresis (2-DE), each REF/SRPP protein was separated into multiple protein spots on 2-DE gels, indicating they have multiple protein species. The abundance of REF/SRPP proteins was compared between ethylene and control treatments or among rubber tree clones with different levels of latex productivity by analyzing 2-DE gels. The total abundance of each REF/SRPP protein decreased or changed a little upon ethylene stimulation, whereas the abundance of

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multiple protein species of the same REF/SRPP changed diversely. Among the three rubber tree clones, the abundance of the protein species also differed significantly. Especially, two protein species of REF175 or REF258 were ethylene-responsive only in the high latex productivity clone RY 8-79 instead of in RY 7-33-97 and PR 107. Some individual protein species were positively related to ethylene stimulation and latex productivity. Distinct functions of the primary and secondary laticifer cells in the rubber tree Tan Deguan, Hu Xiaowen, Fu Lili, Kumpeangkeaw Anuwat, Ding Zehong, Sun Xuepiao, Zhang Jiaming. (ITBB-CATAS, ZES-CATAS, Song Khla Rubber Research) Morphological comparison using paraffin sections revealed only distribution differences: the primary laticifers were distributed randomly, while the secondary laticifers were distributed in concentric rings. Using isolated laticifer networks, the primary laticifers were shown to develop via intrusive "budding" and formed necklace-like morphology, while the secondary laticifers developed straight and smooth cell walls. Comparative transcriptome analysis indicated that genes involved in cell wall modification, such as pectin esterase, lignin metabolic enzymes, and expansins, were highly up-regulated in the primary laticifers and correspond to its necklace-like morphology. Genes involved in defense against biotic stresses and rubber biosynthesis were highly up-regulated in the primary laticifers, whereas genes involved in abiotic stresses and dormancy were up-regulated in the secondary laticifers, suggesting that the primary laticifers are more adequately prepared to defend against biotic stresses, while the secondary laticifers are more adequately prepared to defend against abiotic stresses. Therefore, the two types of laticifers are morphologically and functionally distinct. Effect of HbDHN1 and HbDHN2 genes on abiotic stress responses Cao Yunxin, Xiang Xian, Geng Mengting, You Qin, Huang Xi (Hainan University) HbDHN1 and HbDHN2 expression was examined under multiple abiotic stresses; namely, cold, salt, drought, wounding, abscisic acid (ABA), ethylene (ET), and jasmonic acid (JA) treatment. Although, both HbDHNs were defined as SK2-type dehydrin, they showed different cellular localizations. Overexpression of the HbDHNs in Arabidopsis thaliana further revealed a significant increase in tolerance to salt, drought and osmotic stresses. Increased accumulation of proline and a reduction in electrolyte leakage were also observed under salt and drought stress, and a higher water content was indicated under osmotic stress. The transgenic plants also showed higher activity levels of ascorbate peroxidase (APX), superoxide dismutase (SOD) and catalase, and accumulated less hydrogen peroxide (H2O2) and superoxide. HbICE1 functions in cold tolerance Yuan Hong-Mei, Sheng Ying, Chen Wei-Jie, Lu Yu-Qing, Tang Xiao, Ou-Yang Mo, Huang Xi. (Hainan University, Wuhan University) A nucleic protein with typical features of ICEs, HbICE1 was able to bind to MYC recognition sites and had strong transactivation activity. HbICE1 was constitutively expressed in all tested tissues, with highest levels in the bark, and was up-regulated when subjected to various stresses including cold, dehydration, salinity and wounding. When overexpressed in Arabidopsis, 35S::HbICE1 plants showed enhanced cold resistance with increased proline content, reduced malondialdehyde (MDA) metabolism and electrolyte leakage, and decreased reactive oxygen species (ROS) accumulation. Expression of the cold responsive genes (COR15A, COR47, RD29A, and KIN1) was also significantly promoted in 35S::HbICE1 compared to wild-type plants under cold stress. Differentially expressed genes (DEGs) analysis showed that cold treatment changed genes expression profiles involved in many biological processes and phytohormones perception and transduction. Ethylene, JA, ABA, as well as ICE-CBF signaling pathways might work synergistically to cope with cold tolerance in rubber tree.

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Identification of genes related to rubber formation in Taraxacum koksaghyz Rodin Cao Xin-Wen, Yan Jie, Lei Ji-Liang, Li Jin, Zhu Jian-bo, Zhang Hui-Yan (University of Shi He Zi) The Taraxacum koksaghyz Rodin (TKS) root transcriptome after wild TKS seedlings were sequenced by the treatment of methyl jasmonate (MeJA) for 0, 6, and 24 h. The clean reads generated for each experimental line were assembled into 127,833 unigenes. The Kyoto encyclopedia of genes and genomes pathway prediction suggested that methyl jasmonate regulated secondary metabolism in TKS. Differential expression analysis showed that the expression of HMGCR, FPPS, IDI, GGPPS, and REF/SRPP increased with methyl jasmonate treatment. Interestingly, differential expression analysis of the jasmonate (JA)-related transcription factors (TFs), indicated that certain genes encoding these transcription factors (namely, bHLH, MYB, AP2/EREBP, and WRKY) showed the same expression pattern in the lines treated for 6 h and 24 h. Moreover, HMGCR was up-regulated in the transgenic seedlings overexpressing DREB. Transcriptome-wide identification and characterization of MYB transcription factor genes in the laticifer cells of Hevea brasiliensis Wang Ying, Zhan Di-Feng, Li Hui-Liang, Guo Dong, Zhu Jia-Hong1, Peng Shi-Qing (ITBB-CATAS, Hainan University) 44 MYB genes (named HblMYB1 to HblMYB44) were identified by using our previously obtained transcriptome database of rubber tree laticifer cells and the public rubber tree genome database. Expression profiles showed that five MYB genes were highly expressed in the laticifers. HblMYB19 and HblMYB44 were selected for further study. HblMYB19 and HblMYB44 bound the promoters of HbFDPS1, HbSRPP, and HRT1 in yeast. Furthermore, the transient overexpression of HblMYB19 and HblMYB44 in tobacco plants significantly increased the activity of the promoters of HbFDPS1, HbSRPP, and HRT1. List of publications - CHINA Cao Y, Xiang X, Geng M, You Q, Huang X (2017) Effect of HbDHN1 and HbDHN2 genes on abiotic stress responses in Arabidopsis. Front Plant Sci 8:470. Cao X, Yan J, Lei J, Li J, Zhu J, Zhang H (2017) De novo Transcriptome sequencing of MeJA-induced Taraxacum koksaghyz Rodin to identify genes related to rubber formation. Sci Rep 7:15697. Chao J, Yang S, Chen Y, Tian W-M (2017) Transcript profiling of Hevea brasiliensis during latex flow. Front Plant Sci 8:1904. Guo D, Zhou Y, Li H-L, Zhu J-H, Wang Y, Chen X-T, Peng S-Q (2017) Identification and characterization of the abscisic acid (ABA) receptor gene family and its expression in response to hormones in the rubber tree. Sci Rep 7:45157. Sui J-L, Xiao X-H, Qi J-Y, Fang Y-J, Tang C-R (2017) The SWEET gene family in Hevea brasiliensis - its evolution and expression compared with four other plant species. FEBS Open Bio 7:1943–1959. Tan D, Hu X, Fu L, Kumpeangkeaw A, Ding Z, Sun X, Zhang J (2017) Comparative morphology and transcriptome analysis reveals distinct functions of the primary and secondary laticifer cells in the rubber tree. Sci Rep 7:3126. Tong Z, Wang D, Sun Y, Yang Q, Meng X, Wang L, Feng W, Li L, Wurtele ES, Wang X (2017) Comparative proteomics of rubber latex revealed multiple protein species of REF/SRPP family respond diversely to ethylene stimulation among different rubber tree clones. Int J Mol Sci 18. Wang Y, Zhan D-F, Li H-L, Guo D, Zhu J-H, Peng S-Q (2017) Transcriptome-wide identification and characterization of MYB transcription factor Genes in the laticifer cells of Hevea brasiliensis. Front Plant Sci 8:1974. Wuyun T-N, Wang L, Liu H, Wang X, Zhang L, Bennetzen JL, Li T, Yang L, Liu P, Du L, Wang L, Huang M, Qing J, Zhu L, Bao W, Li H, Du Q, Zhu J, Yang H, Yang S, Liu H, Yue H, Hu J, Yu G, Tian Y, Liang F, Hu J, Wang D, Gao R, Li D, Du H (2018) The hardy rubber tree genome provides insights into the evolution of polyisoprene biosynthesis. Mol Plant 11:429–442. Yuan H-M, Sheng Y, Chen W-J, Lu Y-Q, Tang X, Ou-Yang M, Huang X (2017) Overexpression of Hevea brasiliensis HbICE1 enhances cold tolerance in Arabidopsis. Front Plant Sci 8:1462.

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Zhao M, Liu H, Deng Z, Chen J, Yang H, Li H, Xia Z, Li D (2017) Molecular cloning and characterization of S-adenosylmethionine decarboxylase gene in rubber tree (Hevea brasiliensis). Physiol Mol Biol Plants 23:281–290. Zou Z, Liu J, Yang L, Xie G (2017) Survey of the rubber tree genome reveals a high number of cysteine protease-encoding genes homologous to Arabidopsis SAG12. PLoS ONE 12:e0171725. Zou Z, Xie G, Yang L (2017) Papain-like cysteine protease encoding genes in rubber (Hevea brasiliensis): comparative genomics, phylogenetic, and transcriptional profiling analysis. Planta 246:999–1018.

INDIA Sequencing and annotation of the whole genome of Hevea brasiliensis Advance Centre for Molecular Biology & Biotechnology Scientists Whole genome sequencing of rubber was performed using Next Generation Sequencing (NGS) platforms to generate a draft sequence of rubber clone RRII 105. Sequence data generation (470 GB) were completed with four sequencing platforms (Illumina HiSeq, MiSeq Roche GS-FLX and PacBio). Preliminary de novo genome assembly was performed successfully using MaSuRCA assembler using limited sequence dataset. Around 250 Gb of HiSeq and other mate-pair libraries 4, 6, 8 & 20 Kb) were not included in this assembly. Unfortunately the genome assembly process using MaSuRCA was stopped with complete set of sequence data. Therefore, further stringent filtration was performed on entire sequence data, especially on mate-pair (MP) data to reduce the data size for assembly. Simultaneously, we could do benchmarking for de novo assembly of rubber genome using two assemblers on IBM POWER processor to assess the performance of the server after getting remote access to the same. Hevea transcriptomics Advance Centre for Molecular Biology & Biotechnology Scientists Bioinfomatic analysis of 15 transcriptomes derived from leaf, root, bark and latex of rubber plant is in progress. Protein domain detection was carried out for all 15 RNA-Seq assemblies with Pfam database, which would help in functional annotation of the transcripts. Highest Pfam hit was obtained with bark transcriptome RRI-BA (34,398) and lowest hit with TPD affected bark transcriptome T1 (7,990). Identification of tissue specific transcripts, further annotation and screening of top 50 differentially expressed genes from RNA-Seq projects have been initiated. Root-specific transcripts from root transcriptome of rubber T. Saha and Anantharamanan R. To identify root specific transcript sequences using RNA-Seq assembly, leaf, latex and bark transcript sequences were clustered with 90% sequence identity. A total of 38,047 root transcript sequences were subjected to BLAST and FASTA alignment search against the clustered leaf, latex and bark sequences. A total of 344 root transcripts, which are not having any significant alignment with other tissues were extracted using in-house PERL scripts. Further, these 344 sequences were subjected to Open Reading Frames (ORFs) search, which resulted in 38 transcript sequences with full-length ORFs. Analysis of abiotic stress-responsive transcripts of Hevea brasiliensis M.B. Mohamed Sathik, Lisha P. Luke, Anantharamanan R. et al. Transcriptome sequencing was performed in leaf samples of H. brasiliensis exposed to drought as well as cold stresses using Illumina sequencing technology (RNA-Seq) in order to generate functional genomic resource data which might eventually provide details on molecular mechanisms underlying drought/cold responses in H. brasiliensis. Annotated transcriptome data of both drought and cold stress-responsive transcripts were analyzed and further validated by quantitative gene expression analyses. The digital gene expression analysis indicated an up-regulation of 268 transcripts and down-regulation of 566 transcripts under drought stress while 961 and 109 transcripts were found up- and down regulated respectively, under cold stress. Quantitative gene expression analysis of selected drought responsive transcripts revealed the association between transcripts such as ferritin, DNA-

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binding protein, NAC tf and aquaporin with drought tolerance, and ethylene responsive transcription factor (ERF) with cold tolerance. SNPs in rubber biosynthesis genes and their haplotypes Thomas KU and T. Saha Comparison of the SNP number and frequency in the eight rubber biosynthesis genes: Upstream genes in MVA pathway like HMGS, HMGR. MVK and PMVK are found to be highly conserved compared to the genes REF and CPT with very high SNP frequency rates, which are functionally specific to Hevea brasiliensis for latex biosynthesis. REF had the highest number of SNPs (32) and also recorded with highest frequency (1SNP/53 nucleotides) as well as highest number of non-synonymous SNPs (15) among eight rubber biosynthesis genes. The initiator molecule synthesising genes viz., FDPS and GGDPS are moderately conserved. Interestingly FDPS and HMGR didn’t harbour any non-synonymous SNPs. FDPS had the highest number of SNPs from non-coding regions. SNP marker development: SNP markers developed from the gene GGDPS (HbGGP741C/T & HbGGP1173A/G), MVK (HbMVK2628A/G, HbMVK3297C/T), PMVK (HbPMVK1786C/T), HMGS (HMGS3059A/G) and CPT (HbCPT760A/G & HbCPT1438C/T) were tested for polymorphisms between the parents (RRII 105 and H. benthamiana) of an interspecific mapping population using HRM genotyping methodology. Only the SNP markers MVK2628A/G, PMVK1786C/T, HMGS3059A/G and cis-pre1438C/T were found polymorphic and used in segregation analysis of the progeny population for its integration in the linkage map. Segregation ratio of MVK2628A/G was found to be approximately 1:1 as expected. For PMVK marker the segregation ratio was highly skewed. SNP markers generated from the HMGS (HMGS3059A/G) and CPT (cis-pre1438C/T) genes were segregated at a ratio of 1:1. Construction of a high-density genetic linkage map in rubber Bindu Roy, Jayasree Madhavan, T. Saha High density linkage map was constructed using DArT SNPs and presence/absence variants (PAVs) markers. Refinement of the linkage maps was carried out eliminating single-dose markers and markers showing excessive segregation distortion. Finally 5,187 and 8,082 good quality markers for H. brasiliensis and H. benthamiana respectively were used in linkage map construction for each parent. Linkage maps of both the parents H. brasiliensis and H. benthamiana assembled into 18 linkage groups, which is in agreement with the known number of haploid chromosomes in Hevea (n = 18). Total length of the map was 6666.2 and 8779.9 cM for H. brasiliensis and H. benthamiana respectively. The length of each linkage group in H. brasiliensis ranged from 175.5 cM (LG 13) to 580.7 cM (LG 10), the distance between two markers was on an average 1.28 cM and the maximum space between two markers was 25.2 cM. Percentage of markers participated in mapping was 85%. In case of H. benthamiana, length of each linkage group ranged from 304 cM (LG 17) to 685 cM (LG 5), distance between two markers was on an average 1.09 cM with a maximum space between two markers 16.2 cM. Percentage of markers that were able to map was 71%. QTL mapping of disease resistance in rubber Bindu Roy, T. Saha et al. The genotypic and phenotypic data for disease resistance from the progeny population were merged together to identify marker trait relationship for identification of QTLs related to resistance against major fungal pathogens of rubber: Phytophthora spp., Corynespora cassiicola and Colletotrichum spp. QTLs were detected using ‘qtl’ software in R package. The logarithm of the odds (LOD) score threshold was estimated by permutation test and a LOD score threshold score of 2 with an adjusted P value cut off 0.001 was used. The most probable position of the QTL and the peak LOD score of the QTL were assessed.

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QTLs for resistance to these three diseases were identified from the linkage map of disease resistant parent H. benthamiana. Three QTLs for Phytophthora disease resistance [two mapped in linkage group (LG) 8 and one in LG 1] and three QTLs for Corynespora disease resistance (one each mapped in LG 1, 12, 13) were identified from H. benthamiana. In case of Colletotrichum, QTLs were identified in both H. brasiliensis and H. benthamiana. Three QTLs (one each mapped in LG 10, 12, 18) from H. benthamiana and three QTLs (one each mapped in LG 5, 6, 11) from H. brasiliensis were identified. Validation of QTL markers and fine mapping of the QTL regions for disease resistance was carried out using KASP marker technology. Genetic assessment of international rubber clones received through clone exchange program T. Saha, KU Thomas, KK Mydin et al. RRII could successfully establish best 40 clones from almost all rubber growing countries: Indonesia, Cambodia, Thailand, Philippines, Vietnam, Sri Lanka, Myanmar, Ghana and Cote D’Ivoire through clone exchange program. Genetic assessment of these clones along with the high yielding clones existing in RRII, especially the 400 series clones and RRII 105 was carried out using molecular markers to know the extent of genetic diversity existing among these clones compared to RRII clones and also to develop clone specific SSR profiles for their identity. So far 16 SSR markers were used in genotyping of 34 clones.

Development of gynogenic haploids through embryo sac culture Sushamakumari S. and Jayashree R. The embryogenic callus and embryos obtained from the embryo sac cells of the unfertilized ovules was sub cultured for embryo induction, maturation and plant regeneration. Embryo sac derived calli of clones RRII 105 and RRII 414, generated in the previous year, were proliferated after which they were transferred to embryo induction media. Embryogenic callus induced in clone RRII 414 gave rise to several embryos. In RRII 105 embryogenesis was achieved at a low frequency. These embryos were sub cultured to maturation medium. Some of the matured embryos of the clone RRII 414 are in the germination phase. Generation of tetraploids/polyploids from diploid callus Sushamakumari S. et al. Fresh embryogenic callus emerging from the colchicine treated callus of clone RRII 105 were proliferated and transferred to embryo induction medium. Embryogenesis from these cultures is awaited. In an earlier experiment calli derived from zygotic explant were subjected to colchicine treatment and were maintained by sub culturing to appropriate media combinations. Embryo induction and maturation followed by embryo germination could be obtained. Many of the germinating embryos have been transferred to the plant regeneration medium. Regenerated plantlets are now in the acclimatisation phase. Somatic embryogenesis from leaf explants Kala RG et al. Callus induced from leaf cultures from clone RRII 105 and 400 series clones was proliferated in medium containing silver nitrate (1 mg/l) and picloram (3 mg/l) and made friable. In another experiment, the effect of 2-chloroethyl phosphonic acid (CEPA) on embryogenic callus induction was studied. Compact callus obtained in callus induction medium with higher cytokinin (2ip/BA) along with auxins (2,4-D/NAA) was maintained in the same medium without subculture. After two - three months, this callus was subcultured in medium supplemented with different concentrations (0-5 mg/l) of an ethylene inducer, 2-chloro ethyl phosphonic acid (CEPA). Embryogenic callus induction was obtained from the compact callus after about one month, in medium with CEPA (2mg/l). Somatic embryo induction from the proliferated embryogenic callus was obtained in earlier standardized medium. Embryos after initial one week culture in maturation medium germinated after slow desiccation. In vitro plant regeneration was obtained in earlier standardized medium.

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Genetic transformation of Hevea brasiliensis with MnSOD gene construct with CaMV35S promoter Jayashree R, Kumari Jayasree, Kala RG et al. Agrobacterium-mediated genetic transformation experiments was carried out with HbMnSOD gene construct using embryogenic callus derived from anther, ovule and leaf explants of Hevea clone RRII 105. Several antibiotic resistant cell lines that emerged from the infected target tissues were selected after performing GUS assay and the cell lines were proliferated and tried for embryo induction. The transgenic lines obtained last year failed to produce embryos and therefore attempts were also made to develop new transgenic lines integrated with two genes namely, hmgr1 and MnSOD. Transformation experiment was done with the HbMnSOD gene construct using hmgr1 transgenic cell line as the initial explant. The new transgenic cell lines emerging in the selection medium were transferred for proliferation. Agrobacterium-mediated transformation of Hevea brasiliensis with sorbitol-6-phosphate dehydrogenase (S6PDH) gene Kumari Jayasree et al. Genetic transformation work with two high yielding clones RRII 414 and RRII 430 was also initiated for increasing the stress tolerance. Agrobacterium-mediated genetic transformation was carried out with proliferated friable callus obtained from immature anthers for incorporation of S6PDH genes under the control of CaMV 35S promoter. Viable tissues obtained after bacterial infection were selected and cultured on selection medium for the production of transformed cell lines. Agrobacterium-mediated transformation with the ipt gene Kala RG et al. The plant regeneration system from leaf explants was utilized for Agrobacterium mediated genetic transformation for incorporation of iso-pentenyltransferase (ipt) gene. Work was executed transgenic tissue regeneration, embryo induction and transgenic plant regeneration. Tissue infection were carried out thrice with the ipt gene using proliferated friable fresh callus and embryogenic callus obtained from leaf explants of clone RRII 105 and RRII 414 as target tissues. Several putatively transgenic lines were obtained. A few lines from these were proliferated and somatic embryo induction obtained from two lines incorporated with the ipt gene. Agrobacterium-mediated transformation with the hmgr1 gene Jayashree R. et al. Transgenic plants integrated with hmgr1 were developed and hardened earlier. DNA was extracted from the leaves of these plants. PCR analysis was carried out to monitor the presence of the integrated hmgr1 gene in the plants. Positive amplification for the hmgr1 transgene was observed in twenty out of 36 plants tested and 13 plants were hpt positive. Plants attaining a girth of 15 to 20 cm were tapped and the rubber yield was recorded under containment facility. The transgenic plants showed a range of latex yield values where some plants produced more latex than the control plant indicating possible regulatory role of hmgr1 gene in the latex biosynthetic pathway. Transformation using the callus derived from immature ovules produced 40 transgenic cell lines and 12 cell lines were proliferated and cultured for somatic embryogenesis. Embryogenesis was noticed in four transgenic cell lines and five transgenic plants were developed. Development of transgenic plants integrated with osmotin gene Rekha K. et al. Agrobacterium infections were carried out with clonal as well as zygotic explants for developing new transgenic events integrated with osmotin gene derived from tobacco plant. Transgenic lines were developed and proliferated. Mature embryos were transferred to new media combinations for improving the regeneration percentage as well as hardening. About 10 transgenic plants were regenerated in the new media combination and transferred for hardening. Out of them, 3 plants survived the initial phase of hardening and survived until 3 months. However they failed to develop

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further. Experiments are continuing for developing new plants. For the evaluation of drought tolerance in earlier developed plants, the membrane stability was assessed using the standard protocol. Transgenic plants showed better membrane stability, compared to control, which is an indication of the positive expression of inserted gene towards stress tolerance. List of publications – INDIA Kala R.G., Supriya R., Mathew S.A., Jayashree R. and Thulaseedharan A. (2017). Cloning and characterization of a full-length CAB gene encoding the light harvesting Chlorophyll A/B Binding Protein In Hevea brasiliensis. Rubber Science, 30(1): 25-41 Kumari Jayasree P., Saseedhran S. and Mathew S.A. (2017). Embryogenic competence and HbSERK gene expression during somatic embryogenesis, Rubber Science, 30(1): 76-81 Mohamed Sathik M.B., Luke L.P., Rajamani A. et al. (2018). De novo transcriptome analysis of abiotic stress-responsive transcripts of Hevea brasiliensis. Mol Breeding (2018) 38: 32. https://doi.org/10.1007/s11032-018-0782-5 Jacob J., Jayashree R., Thulaseedharan A. and Kala R.G. (2017). Hevea SOD transgenics - way to the field. Fifth Annual South Asia Biosafety Conference, September 11-13, Taj West End, Bangalore. Jayashree R., Venkatachalam P., Kala R.G., Leda P. and Nazeem P.A. (2017) Hmgr1 transgene integrated Hevea - a future candidate to the field. Fifth Annual South Asia Biosafety Conference, September 11-13, Taj West End, Bangalore. Thomas K.U. and Saha T. (2017) SNP identification and haplotype structuring in the latex biosynthesis genes of Hevea brasiliensis. CATAS/ IRRDB Workshop on Rubber Genomics and Molecular Genetics 2017, Haikou, Hainan, China, 27-29 September 2017. Rajamani A., Mohamed Sathik M.B. and Thomas M. (2017) Comprehensive assessment of RNA-Seq mapping strategies for measuring differential gene expression. CATAS/ IRRDB Workshop on Rubber Genomics and Molecular Genetics 2017, Haikou, Hainan, China, 27-29 September 2017. Roy C.B., Killian A., Madhavan J. and Saha T. (2017) Genetic dissection of disease resistant trait in rubber (Hevea brasiliensis) through QTL mapping. International Rubber Conference & IRRDB Annual Meetings 2017: Current and Future Challenges in Natural Rubber Industry: Capturing Opportunities in The Green Economy. Jakarta, Indonesia, 18 – 20 October 2017. Rajitha.K.P., Philip S., Sushamakumari S. (2017) Preliminary investigations on Corynespora leaf fall disease of Hevea brasiliensis through in vitro screening and biochemical assay, Proceedings of Kerala Environment Congress held at Energy Management Centre, Thiruvananthapuram , 6 - 8 December, p-207.

FRANCE Genomic and transcriptomic analyses Julie Leclercq, Yi Zhang, Shuangyang Wu, Bénédicte Favreau, Pascal Montoro A deep RNA sequencing of latex has been carried out to compare the effect of ethephon stimulation and Tapping Panel Dryness (TPD) severity. Differentially expressed genes were identified and annotated, giving 8,111 and 728 in response to ethephon in slight TPD trees and in ethephon-induced severe TPD trees, respectively. A biological network of responses to ethephon and TPD highlighted the major influence of metabolic processes and the response to stimulus, especially wounding and jasmonate depression in TPD-affected trees induced by ethephon stimulation [1]. A genome-wide analysis of redox-related genes has been carried out using genome sequence published by Tang et al. (2016) and RNAseq data (see above). The Hevea genome harbours 407 genes of the thirty main redox-related gene families. There are 161 redox-related genes expressed in latex. Thirteen of these redox-related genes were targeted by 12 microRNAs and 15 genes by 16 phased siRNA [2, 3]. In response to environmental cues and latex harvesting stress, Hevea revealed an alteration in sRNA transcriptome in latex cells associated with physiological disorders halting latex flow (Tapping Panel Dryness, TPD). Most sRNAs are 24 nt in healthy trees and 21 nt in TPD-affected trees. This questions

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the contribution of post-transcriptional gene silencing by sRNA to TPD. Degradome data were used to highlight sRNA-mediated post-transcriptional genome expression regulation in response to abiotic stress and TPD. We sequenced all 3’ ends of degraded mRNAs from 6 tissues, subjected or not to abiotic stress/TPD, to validate all sRNA-based target cleavage sites simultaneously. Latex cells form a tissue with the highest level of post-transcriptional regulation by mRNA cleavage and natural rubber (NR) biosynthetic pathway is under a strong post-transcriptional silencing. Partial conservation of miRNA-mediated post-transcriptional regulation was observed between Hevea and Arabidopsis [4, 5]. List of publications - FRANCE 1Montoro P, Wu S, Favreau B, Herlinawati E, Balzergue S, Magniette M-L, Pointet S, Rio M, Leclercq J, Ismawanto S et al (2018). Transcriptome analysis in Hevea brasiliensis latex revealed changes in hormone signalling pathways in response to ethephon stimulation and Tapping Panel Dryness occurrence. Sci Rep 8:8483-8494. 2Zhang Y, Leclercq J, Montoro P (2017). Reactive oxygen species in Hevea brasiliensis latex and relevance to Tapping Panel Dryness. Tree Physiol 37(2):261-269. 3Zhang Y, Leclercq J, Wu S, Ortega-Abboud E, Pointet S, Tang C, Hu S, Montoro P (submitted). Genome-wide analysis of genes involved in production and scavenging of reactive oxygen species and antioxidant biosynthesis in Hevea brasiliensis laticifers. 4Leclercq J, Wu S, Farinas B, Pointet S, Favreau B, Vignes H, Kuswanhadi, Dufayard J-F, Gao S, Hu S et al (submitted). Post-transcriptional regulation of rubber production in Hevea brasiliensis laticifers. 5 Leclercq J, Wu S, Montoro P (2017). Towards genome-wide analysis of transcriptional and post-transcriptional regulations in Hevea. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China.

JAPAN Improvement of sequence of the Hevea brasiliensis RRIM 600 genome Yuko Makita, Mika Kawashima, Minami Matsui in collaboration with Keng-See Chow, MRB Not much progress has been made in the improvement of RRIM 600 draft genome sequence. Effort was put into assembling program using long reads. We also performed Hi-C library construction of RRIM 600 clones in collaboration with MRB, Malaysia. In vitro reconstruction of rubber biosynthesis Satoshi Yamashita, Haruhiko Yamaguchi, et al. To aim at reconstruction of rubber biosynthesis system, an in vitro synthesis of natural rubber was developed in Tohoku University in collaboration with Sumitomo Rubber Industries Ltd. Ref: Satoshi Yamashita, Haruhiko Yamaguchi, Toshiyuki Waki, Yuichi Aoki, Makie Mizuno, Fumihiro Yanbe, Tomoki Ishii, Ayuta Funaki, Yuzuru Tozawa, Yukino Miyagi-Inoue, Kazuhisa Fushihara, Toru Nakayama and Seiji Takahashi (2016). Identification and reconstitution of the rubber biosynthetic machinery on rubber particles from Hevea brasiliensis. eLife 2016;5:e19022. Genome browser for transcriptome including CAGE data and full-length cDNAs information Yuko Makita, Mika Kawashima, Lau Nyok Sean, Ahmad Othman Sofiman, Minami Matsui In collaboration between RIKEN and Universiti Sains Malaysia, a transcriptome database was constructed based on the RRIM 600 genome browser. This browser includes information of full-length cDNAs, CAGE (Cap-analysis Gene expression data) and RNA-Seq data of gene expression produced in-house and from public data. The Rubber Transcriptome DB release 2017.03 is accessible at http://matsui-lab.riken.jp/rubber/. Other domestic activities Bridgestone reported rubber genome sequence improvement during the 35th JSPMB conference in 2017, entitled “Improvement of Genome Sequence of a Rubber tree Hevea brasiliensis”.

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List of publications – JAPAN: Yuko Makita, Mika Kawashima, Nyok Sean Lau, Ahmad Sofiman Othman and Minami Matsui (2017). Construction of Pará rubber tree genome and multi-transcriptome database accelerates rubber researches. 16th InCOB, Shenzhen, China. Published in BMC Genomics 19 (Suppl. 1):922 on 19 Jan 2018 (https://doi.org/10.1186/s12864-017-4333-y). Minami Matsui (2017). Rubber genome and CAGE analysis of its transcriptome. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China. THAILAND Transcriptome analysis of ethylene response in Hevea brasiliensis Piyanuch Piyatrakul and Sithichoke Tangphatsornruang (RAOT/NSTDA) The use of latex accelerators such as Ethephon, an ethylene release agent, has been widely used to increase the amount of latex yield. The use of latex accelerator chemicals can increase the flow time of the latex up to twice or more. Ethylene also promotes the metabolism of latex cells and sucrose transport, which is the precursor to the formation of rubber particles from neighboring cells into the latex cell. However, ethylene reaction of latex yield also depends on the genetic. The influences of ethylene that affect the yield of latex are unclear. Thus, the transcriptome of rubber under ethylene-stimulated conditions were studied by comparing between normal condition in high yielding rubber variety (RRIT 251) and low yielding rubber variety (AVROS2037). The results can lead to the search for relevant genes. Including understanding the mechanism of gene expression. The result can be applied as a marker assisted selection (MAS) with precision as it is a direct marker or functional marker assisted selection (fMAS) to optimize and shorten the duration of conventional breeding. Genome-wide association mapping of latex yield and girth in Amazonian accessions of Hevea brasiliensis grown in a suboptimal climate zone Vipavee Chanroj, Ratchanee Rattanawong, Thitaporn Phumichai, Sithichoke Tangphatsornruang and Kittipat Ukoskit (TU/RAOT/NSTDA) Latex yield and growth are the key complex traits in commercial rubber production. The present study is the first to report genome-wide association mapping of latex yield and girth, for 170 Amazonian accessions grown in a suboptimal area characterized by limited rainfall and a lengthy dry season. Targeted sequence enrichment to capture gene transcripts generated 14,155 high quality filtered single nucleotide polymorphisms (SNPs) of which 94.3% resided in coding regions. The rapid decay of linkage disequilibrium over physical and genetic distance found in the accessions was comparable to those previously reported for several outcrossing species. A mixed linear model detected three significant SNPs in three candidate genes involved in plant adaptation to drought stress, individually explaining 12.7–15.7% of the phenotypic variance. The SNPs identified in the study will help to extend understanding, and to support genetic improvement of rubber trees grown in drought-affected regions. Development of molecular marker associated to Hevea leaf disease Kannikar Teerawattanasuk and Sithichoke Tangphatsornruang (RAOT/NSTDA) Hevea brasiliensis, commonly known as the rubber tree, is almost the sole source of natural rubber production. Natural rubber has a wide range of industrial applications and is under increasing global demand. Rubber tree is the most widely cultivated tree species for producing natural rubber latex. Research programs have focused on increasing latex yield and responses to abiotic and biotic stresses. The research team sequenced the genome of the rubber tree BPM24 variety which exhibits a high degree of disease resistance to several pathogens and also shows cytoplasmic male sterility. We reported the rubber tree genome as a reference sequence from de novo assembly (PacBio, Illumina, 454, and Chicago libraries) and mutations on the mitochondrial genome that cause cytoplasmic male sterility in BPM24. RNA sequencing from several rubber tree varieties have been performed to identify SSR and SNP markers. High throughput genotyping techniques based on Target Enrichment and

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Reduced Representation Libraries have developed to construct a combined genetic linkage map from several populations. Linkage analysis on mapping populations and germplasm have been performed to identify markers related with latex yield and disease resistance. List of publications – THAILAND: Chanroj V., Rattanawong R., Phumichai T., Tangphatsornruang S., Ukoskit K. (2017). Genome-wide association mapping of latex yield and girth in Amazonian accessions of Hevea brasiliensis grown in a suboptimal climate zone. Genomics 109 (5-6): 475-484 Pootakham W., Sonthirod C., Naktang C., Ruang-Areerate, P., Yoocha, T., Sangsrakru, D., Theerawattanasuk, K., Rattanawong, R., Lekawipat N., Tangphatsornruang S. (2017). De novo hybrid assembly of the rubber tree genome reveals evidence of paleotetraploidy in Hevea species. Sci Rep 6, 41457. Phumichai T. (2017). Genome-wide association studies (GWAS) in Hevea. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China.

INDONESIA Genetic and genomic analysis of Tapping Panel Dryness (Collaborative research project between Indonesia Rubber Research Institute and CIRAD) Fetrina Oktavia, Eva Herlinawati, Martini Aji, Sigit Ismawanto, Kuswanhadi, Maryannick Rio, Julie Leclercq, Vincent Le Guen, Andre Clement-Demange, Regis Lacote, Pascal Montoro Tapping Panel Dryness (TPD) occurrence is one of the major agronomical issues affecting latex production. Molecular mechanism involved in TPD occurrence has been intensively studied and many candidate genes have been identified. However, no information has been published on the allelic variability of these factors in relation with TPD occurrence. Recent molecular physiology studies suggested a specific phenotypic methodology to study genetic inheritance of TPD tolerance or susceptibility. 201 progenies from susceptible clone PB 260 and tolerant clone SP 217 has been planted in the field in December 2016. Two parental clones (PB 260 and SP 217) and five recommended clones i.e. AVROS 2037, GT 1, IRR 112, IRR 39, PR 261 and RRIC 100 were used as controls. Legitimacy test was carried out using 5 SSR markers to verify the progenies planted in the field. Based on the legitimacy test all progenies planted in the field were correct. The plants growth was observed monthly. The growth data collected from 1 year old plants showed that the stem diameter of the progenies ranged from 23.3 mm to 48.67 mm. Moreover, 128 progenies have higher stem diameter than control RRIC 100 (40.94 mm). Genetic diversity analysis of Hevea brasiliensis accessions from Rondonia wild population conserved in Vietnam (RRIV-CIRAD collaboration) PhD Project by Vu Van Truong with Nong Lam University (Supervisors: Dr. Vincent Le Guen, CIRAD and Dr. Huynh Van Biet, Nong Lam University) In the current study, we characterize with SSR markers more than one thousand accessions originating from Rondônia and the ex-situ germplasm collection of RRIV in Vietnam. The RRIV received from Malaysia between 1984 and 1987 a duplication of almost 3,000 IRRDB accessions. These were carefully assessed for their agronomic traits, and characterized with molecular markers. The agronomic evaluation identified several accessions from Rondônia for their rapid growth and relatively good latex production under Vietnamese conditions of cultivation; therefore, they could be worthwhile for both timber and rubber production. The objectives are to establish the real link of each putative Rondônia accession with one of the genetic Rondônia clusters, to check the possible existence of duplication identified under different accession names, and to describe the genetic structure existing within accessions from Rondônia. We will also examine under which conditions it should be possible to conduct a Rondônia-based genome wide-association study for growth and latex production, two agronomic traits of major importance in rubber tree.

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List of publications - INDONESIA Irfan Martiansyah, Riza Arief Putranto and Nurul Khumaida (2017). Identification of putative gene family encoding protease inhibitors by in silico comparative analysis in Hevea brasiliensis Muell. Arg. genome. Menara Perkebunan 85 (2): 53-66. Riza Arief Putranto, Irfan Martiansyah, RT Saptari (2017). In silico identification and comparative analysis of Hevea brasiliensis COBRA gene family. International Conference on Science and Engineering, 39-47. Eva Herlinawati and Sigit Ismawanto (2017). Physiological characters and production toward tapping panel dryness of PB 260. Proceedings of IRRDB International Rubber Conference, 801-810. Fetrina Oktavia (2017). Analysis of molecular markers linked to Corynespora leaf fall disease resistance in rubber plants. Proceedings of IRRDB International Rubber Conference, 170-182.

VIETNAM Biological control of Corynespora cassiicola causing Corynespora leaf fall disease (CLF) on rubber tree by Bacillus thuringiensis (T3) Masters Project by Pham Ngoc Giau with Nong Lam University (Supervisor:Dr. Nguyen Bao Quoc) The aims of the present study are (1) to detect the Cas gene in 20 Corynespora cassiicola isolates (2) to investigate the inhibitory activity of B. thuringiensis in Hevea brasiliensis against Corynespora cassiicola and (3) to evaluate the expression of Cas gene under the induction of the antifungal bacteria B. thuringiensis. Two of the twenty C. cassiicola isolates were classified as Cas2. The other 18 isolates in which no Cas gen was detected were classified as Cas0. Semi-quantitive RT-PCR and Realtime PCR were used to detect gene expression changes in response to antifungal bacteria. The results indicated that cassiicolin gene expression was down-regulated under treatment with B. thuringiensis T3. MALAYSIA Investigating the cassiicolin gene in Malaysian Corynespora cassiicola isolates (Murnita Mohamad Mahyudin) This study was undertaken to investigate the presence of cassiicolin gene in C. cassiicola isolates from Malaysia, emphasizing on isolate C7. Thirteen isolates were selected and subjected to DNA extractions and PCR amplifications using C. cassiicola beta tubulin primers. PCR results showed that DNA extractions were successful and all DNAs were genuinely of C. cassiicola. PCR amplifications were performed on all isolates using cas1, cas2, cas3, cas4, cas5 and cas6 primers to amplify six different cassiicolin isoforms. The PCR results showed that that only isolate C7, C6811, CTD6 and C6810 produced the expected product size of 750 bp, when amplified using cas5 primers. The unpurified PCR products from isolates C7, C6811, CTD6 and C6810 were sent for sequencing and aligned using cas5 sequence from the NCBI database. It was revealed that all the nucleotide sequences were identical to the cas5 gene. The sequences were also translated into six amino acid reading frames and compared with that of cas5 isoform. It was confirmed that all isolates carry cas5 gene in their genome. Pathogenicity assays were performed to evaluate the virulence of cas5-containing isolates. ANOVA results showed that clonal susceptibility to C. cassiicola is highly and significantly affected by clones and isolates (P<0.05); the interaction between isolates and clones used was also significant (p<0.05). Generally, the isolates C7, C6811, CTD6 and C6810 showed light to moderate infection in all clones tested, namely RRIM 600, RRIM 928, RRIM 2020 and PB 260. Sequencing and annotation of a Corynespora cassiicola genome (Murnita Mohamad Mahyudin) The genome of the Malaysian Corynespora cassiicola isolate, C7, was sequenced (as part of a PhD project with University of Bristol, UK: Supervisors: Dr. Andy Bailey and Prof. Gary Foster) and subsequently annotated in MRB. C. cassiicola genome assembly showed that the genome size is 42.5

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Mbp with 774 contigs. Genome Assembly and Gene Set Completeness Assessment showed that the genome is rather complete, with 98.6% conserved orthologs. A total of 13,899 genes was predicted and the total length of predicted genes (intronic regions included) is 21,906,080 bp. The number of predicted protein coding gene (> 33 aa) is 13,527 with a total length of 6,438,937 amino acids. A total of 13,527 peptide sequences (> 33 amino acids) from predicted genes were searched against the NCBI non-redundant and Swissprot database. Approximately 89% and 58% sequences were annotated using NCBI non-redundant and Swissprot, respectively. From the NCBI non-redundant database, the top five species hits were all pathogenic fungi with the highest number of hits to Phaeosphaeria nodorum. Over 8,732 sequences (64.5%) were annotated with 11,454 Gene Ontology (GO) identity and 3,077 predicted proteins corresponded to 1,030 enzymes involving 142 KEGG metabolic pathways. A total of 4,266 genes were identified as putative pathogen-host interaction genes from blast search against the PHI (Pathogen-Host Interaction) database. Genes conferring “reduced virulence” and “unaffected pathogenicity” formed the highest percentage of total hits. Lateral flow diagnostic kit for detection of latex allergens (Alessandra Di Cola, Roslinda Sajari, Norazreen Abd Rahman) A latex allergen diagnostic assay kit named as Latex-T has been developed by a joint effort between researchers at the Tun Abdul Razak Research Centre (TARRC), UK and MRB Sungai Buloh. This simple device can monitor two latex allergens (Hev b5 and Hev b13) in natural rubber latex (NRL) products using a lateral-flow devise (LFD) format. The kits used for pre-commercialization trial were produced in 2013 by a manufacturer in United Kingdom. Since two years ago, effort was dedicated to the production of Latex-T using a Biodot equipment at TARRC. Following this, new set of antibodies against the two kit antigens were developed. The antibody development phase for the two allergens has concluded in 2017. Dipstick tests were carried out using a sandwich-format assay. Further work was then carried out on transferring the dipstick assays into the dry lateral flow assays. Different combinations of sample-pad/conjugation-pad membranes and buffers were tested and the conditions for manufacturing LFDs in-house were finally optimized. Studies on the genetic regulation of clonal variation in rubber molecular weight distribution (Keng-See Chow, Roslinda Sajari) Molecular Weight Distribution (MWD) of latex rubber has long been regarded to be genetically controlled based on unimodal and bimodal types produced from GPC (gel permeation chromatography) analysis of clonal latex. However, such previous studies analyzed pooled latex samples from trees of the same clone and from seedling materials. Therefore, there is a need for validation of genetic regulation of MWD by analyzing samples from individual trees with sufficient biological and technical replicates. A selection of 24 clones from an MRB large scale clone trial was selected for establishing their rubber MWD profiles. For each clone, 6 trees were chosen as biological replicates. To study the effect of age on clonal MWD, fresh latex was also collected from 3 clones grown in polybags, including biological replicates. Due to the large number of samples in this project (including technical replicates), a GPC instrument was dedicated to this project. This is important for ensuring reproducibility of GPC runs and for determining a reliable phenotyping assay for clonal rubber MWD. Within the first year of the project a) a protocol for GPC sample prep and run has been optimized and b) GPC analysis has been completed on dried films of latex collected from mature trees of 24 clones and polybag plants of 3 clones. Compilation of molecular weight parameters and MWD profiles of each clone and the analysis of chromatogram runs continued to the end of 2017. Characterization of transgenes and transgene products of genetically-modified Hevea trees from the MRB confined field planting at the Kota Tinggi Research Station, Johor (Siti Shuhada Shuib, E. Sunderasan) Transgenic Hevea including bud-graft generations harbouring glucoronidase (GUS), a recombinant antibody (scFv4715), human protamine 1 (HP1), and human atrial natriuretic factor (HANF) genes were

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planted in the MRB confined field trial (CFT) at the Kota Tinggi Research Station, Johor. These transgenic Hevea plants are part of a long term monitoring and evaluation plan. A PhD project carried out by Siti Shuhada in Universiti Putra Malaysia (Supervisors: Assoc. Prof. Parameswari Namasivayam and Assoc. Prof. Suhaimi Napis) is part of this plan. Leaf and latex samples sourced from selected stands in the CFT will be used for analysis of foreign genes including their expression patterns and characterization of the encoding proteins in accordance to biosafety regulations. While the main investigation is focused on the transgenic foreign proteins, an interim experiment was also initiated with the aim to compare the levels of allergenic latex proteins in both the transgenic and the non-transgenic Hevea trees in the CFT. Performance of panel BO-1 of RRIM 2025, RRIM 600 and GL 1 tissue culture trees (Nor Mayati Che Husin) Tissue culture derived RRIM 2025, a latex-timber clone, has the potential to serve as feedstock for both conventional and forest plantings. Initial RRIM 2025 tissue culture (TC) experiments yielded a total 15 mother plants (MP), from which 210 (V1) bud-grafted plants were obtained. These plants were planted in the Kota Tinggi Research Station, Johor. The performance of virgin basal panel (BO-1) of RRIM 2025 TC derived MP and their bud-grafted generation (V1) trees were tested in comparison to GL 1 TC trees (MP), RRIM 600 TC trees (MP and V1), conventional bud-grafted RRIM 2025 (NB) and control (RRIM 2025 seedling trees). Normal ½S d3 tapping on BO-1 panel was employed in three different yielding periods: low yielding period (LYP), medium yielding period (MYP) and high yielding period (HYP). Growth in terms of clear bole volume (CBV) was also investigated. The results presented in this report were from the first year of evaluation of trees newly opened for tapping. Yield productivity of TC and bud-grafted trees can be classified into two groups (a) the higher performance group represented by RRIM 2025V1, control (RRIM 2025 seedlings trees), RRIM 2025 (NB) (b) the lower performance group represented by RRIM 2025MP, RRIM 600MP, GL 1MP and RRIM 600V1. Future evaluation of latex yield and additional parameters as well as implementation of suitable latex harvesting technologies will contribute more information on the performance of TC rubber trees. List of publications - MALAYSIA Daniel Brown, Mistianne Feeney, Mathin Ahmadi, Chiara Lonoce, Roslinda Sajari, Alessandra di Cola, Lorenzo Frigerio (2017). Subcellular localization and interactions among rubber particle proteins from Hevea brasiliensis. J Exp Bot 68(18): 5045-5055 R. Greenhill, E. Mollison, C. Dowman, L. Johnson1, S.A.M. Arif, B. Clavijo, V. Serkiova, S. Atan, M. Kolesnikova-Allen (2017). Utility of single nucleotide polymorphism markers in clonal profiling of Hevea brasiliensis. Acta Hortic 1152. ISHS 2017. DOI 10.17660/ActaHortic.2017.1152.58. Proc. III International Conference on Agricultural and Food Engineering (Eds.: S. Khairunniza Bejo et al.) Norazreen Abd-Rahman and Mohd-Fazri Kamaruddin (2017). Comparative proteomic analysis of Hevea latex serum induced by ethephon stimulation. 27th Malaysian Society of Plant Physiology Conference, 21-23 August 2017, Pulai Springs Resort, Johor Nor Mayati, C.H., Izilawati, A. M. (2017). Regeneration of latex timber clone RRIM 3001 via somatic embryogenesis. Trans. Malaysian Soc. Plant Physiol. (ISBN 978-967-10840-6-9) 24:143-146. Nor Mayati Che Husin, Abd Razak Ahmad, Saemah Jabar (2017). Latex yield profile and growth performance of field planted Hevea brasiliensis tissue culture clones. 27th Malaysian Society of Plant Physiology Conference, 21-23 August 2017, Pulai Springs Resort, Johor Keng-See Chow (2017). The Rubber DNA Blueprint: Technological Leverage for the Future. Global Rubber Conference, 12-14 September 2017, Istana Hotel, Kuala Lumpur. Keng-See Chow, Roslinda Sajari and Alessandra di Cola (2017). Rubber biosynthesis in Hevea brasiliensis: journeying from conventional to omics approaches. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China.

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Roslinda Sajari, Azlina Bahari and Keng-See Chow (2017). Molecular characterization of the 1-deoxy-D-xylulose-5-phosphate synthase (DXS) from Hevea brasiliensis. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China. Safiah Atan, Alessandra Di Cola and Nurul Siddiqui (2017). Gene expression and protein changes in Hevea brasiliensis response to infection by soil fungus Rigidoporus microporus. CATAS IRRDB Workshop on Rubber Genomics and Molecular Genetics, 27-29 September 2017, Marriott Hotel, Haikou, China. Roslinda Sajari, Alessandra di Cola, Mok Kok Lang (2017). Latex-T: An Allergen Test Kit. Malaysian Rubber Technology Digest 17(1). Alessandra di Cola and Roslinda Sajari (2017). Detecting Latex Protein Allergens with Latex-T. STRETCH Vol. 11: 30 Keng-See Chow (2017). Wickham Meets Sanger. Malaysian Rubber Technology Digest 17 17(1).

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Report on Socioeconomic Activities of the Specialist Group by

Wasana Wijesuriya, Liaison Officer Activities in 2017/2018 Summary: Research and development activities focused on socioeconomics in the member countries have been mainly focused on retaining the rubber farmers in the rubber plantation sector. Hence the research and development programmes have been targeted to find ways for addressing the low price situation and high cost of production (COP), improving the land productivity, addressing worker shortage and thereby to improve the income status of rubber farmers. Promotion of rubber cultivation in new areas has also given priority. With respect to the low price situation, efficient in-country marketing systems, improvement in the quality of raw rubber to assure a fair price and assessments on incentives and subsidies have been focused. Studies on low intensity harvesting systems have been promoted to reduce COP and to address worker shortage for latex harvesting. Strategic technology transfer methods also have been proposed to improve awareness and subsequently increase adoption to improve productivity of rubber land. Country wise activities: Five member institutions have reported on the socioeconomic activities in their countries. The members of the socio-economic specialist group who reported their activities are given below.

Member Country Representative Institute email

China Prof. Liu Ruijin CATAS liuruijincl@gmail.com

Côte d'Ivoire Dr. Mahayao Adolphe

Centre National de Recherche Agronomique (CNRA)

mahyaoadolphe@yahoo.fr

Indonesia

Ms. Dwi Shinta Agustina

Indonesian Rubber Research Institute

dwishinta_sbw@yahoo.com

India Ms. Binni Chandi Rubber Research Institute of India

binni@rubberboard.org.in

Sri Lanka Dr. Mrs. Wasana Wijesuriya

Rubber Research Institute of Sri Lanka

wijesuriyawasana@gmail.com

The following activities have been conducted in the rubber sector by respective countries on socio-economic aspects during 2017/18. Chinese Academy of Tropical Agricultural Sciences (CATAS)

• Studying the effect of continuing low prices on the adoption of technology and production behaviors including tapping, use of chemicals, intercropping with rubber and plantation transformations

• Design an innovation chain for the integration of planting, processing and manufacturing sectors in Hainan Province

• Study on the response of rubber growers to low prices: willingness expression and behavior choice

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• Analysis on the diffusion dilemma of low-frequency tapping technologies in natural rubber: Evidence from rubber farmers in Hainan, China

• Study on the diffusion process of improved rubber variety CATAS (reyan) 73397 in Hainan province

Centre National de Recherche Agronomique (CNRA), Cote d’Ivoire

• Economic evaluation of organic fertilization of seedlings and young rubber plants

Rubber Research Institute of India

• Study on Block Rubber Plantations in Mayurbhanj District of Odisha state, India

• Trends in seasonality of natural rubber production in major producing countries: a disaggregate level analysis

• Incremental economic worth of the first high yielding Indian hybrid clone, RRII 105 over RRIM 600

• An analysis of the activities performed by the Rubber Producers’ Societies and strategies to improve their performance

• A road map for attaining self-reliance in natural rubber production in India by 2030.

• Over-dependence of Indian rubber industry on imported natural rubber: the question of long-term sustainability

• Comparative advantage and export performance of India’s rubber sector: an exploratory analysis

Indonesian Rubber Research Institute

• Land use optimization of rubber plantation with timber crops : A case study on plantation companies in Central Java

• Application of fertilization by spread : An effort to reduce the amount and cost of labor and its influence on the growth of rubber plants

• Effects of organic fertilizer briquette rawa pening peat on the growth of rubber plant rootstock in coco peat medium in root trainers

• Application of Ethylene gas stimulant technology (C2H4) on rubber plants at the low rubber prices

• Low frequency tapping system to overcome the problem of scarcity of tappers

• Economic analysis of delayed open tapping in rubber plantation to overcome low rubber price

Rubber Research Institute of Sri Lanka

• Analysis of driving forces and the spatial variability of productivity in smallholder rubber lands in Sri Lanka

• Analysis of Technical Efficiency and its determinants of Latex harvesting in Dartonfield Estates, Rubber Research Institute of Sri Lanka

• Analysis of Rubber Based Farming Systems in traditional Rubber growing areas Sri Lanka

• Analysis of impact of CESS tax policy changes on raw rubber exports in Sri Lanka

• Worthiness of investing on rainguards in rubber estates: case study on estates owned by the rubber research institute

• Introduction and Establishment of New Fuelwood Growing Models in Selected Lands of Smallholder Rubber Farmers

• Devising and testing of novel technology transfer methods in the smallholder rubber sector

• Establishing Rubber Cultivation Projects for the Voluntary Carbon Market in Eastern and Uva Provinces of Sri Lanka

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• Identification of socioeconomic and cultural constraints for rubber cultivation in Northern Province of Sri Lanka.

• Impact of rubber cultivation on the rural livelihood in non-traditional areas of the country

Activities Conducted in 2018 International Workshop on Statistical Applications in Socio-economic Research, 01 – 02 October 2018 This workshop was attended by 08 member countries; namely, Malaysia, India, Indonesia, Myanmar, China, Cambodia, Thailand and Sri Lanka. A total of 12 foreign participants and 21 local participants representing the Ministry of Plantation Industries, Rubber Development Department and the Rubber Research Institute of Sri Lanka has participated in the workshop. The workshop covered the following areas of statistical methods; viz. Correlation, linear and non-linear regression, Rank and count data analysis and Multivariate statistics, together with practical sessions with hands-on support. Four special topics, viz. Sustainability Valuation, Time series applications in the rubber sector, Survey design and sampling methods for Socio-economic research and Quantifying Causal Maps through Decision Support Systems. International Forum on Current Socio-economic Issues and their Impact on Rubber Production Sector Performance, 03 – 05 October 2018 This forum was attended by 41 participants, including the foreign participants who attended the workshop on Statistics and local participants representing the Ministry of Plantation Industries, Rubber Development Department, Rubber Secretariat, National Institute of Plantation Management, Sri Lanka Council for Agricultural Policy (SLCARP), Smallholder Tea and Rubber Revitalization (STaRR) Project, Planters’ Association of Sri Lanka and the Rubber Research Institute of Sri Lanka. The Chairman, M. N V T A Weragoda welcomed the delegates while Dr. Abdul Aziz, the Secretary General of IRRDB made the introductory remarks on the Forum on Socio-economic Issues and their Impact on Rubber Production Sector Performance. As guest speakers, Dr. Jacob Mathew, IRRDB Fellow and Dr. Suthee Intraskul, Former Chairman, The Expert Group on Establishment of a Regional Rubber Market, IRCO Thailand addressed the audience on their experiences, viz. Participatory Learning and Action Method to assist in community level organization and implementation of development projects by rubber farmers and solving socio-economic problems of rubber growers in Thailand through effective technology transfer mechanisms, respectively. In the two technical sessions, there were 08 presentations from China, India, Indonesia, Thailand, Malaysia, Cambodia and Sri Lanka. The Forum concluded with a field visit to a promising smallholder field and a tour to the Southern part of Sri Lanka, where the participants experience the mangrove caves, the jungle beach and the famous Dutch Fort in Galle. Activities proposed for 2019 1. Compendium on Issues Related to Technical Inefficiency in Smallholder Rubber Farms in IRRDB Member Countries Socio-economic studies, especially the ones that focus economic efficiency are among the essential components in the rubber industry as it provides necessary information for appropriate decision making. In that sense they provide information for policy makers, to develop policies in one hand and also to evaluate the impact of policies implemented so far, on the other hand. Such information ultimately would help the policy makers to implement efficient policies for the betterment of the rubber industry in respective countries. Issues related to economic efficiency in the smallholder rubber sector have been studied in isolation in several rubber growing countries. Yet, a comparison among these issues through a common approach by respective member countries; starting from data collection to publication of a compendium through the most appropriate statistical and economic methods is yet to be introduced

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for the benefit of individual countries as well as for IRRDB in devising future plans for the benefit of the rubber industry. “Technical efficiency” is defined by Koopmans as: “If a producer needs to decrease one of the outputs or increase one of the inputs in order to increase its output, the situation is technical efficient”. As can be seen in the definition technical efficiency can be analyzed by using input-oriented or output-oriented approaches. The key word focused in this proposal is the “Technical inefficiency”, the inability of a farm to achieve the optimum efficiency, which may be due to technical, socio-economic, environmental and institutional issues. Therefore, the objective of this proposal is to compile a document on the issue of technical inefficiency and related issues in the smallholder rubber sector of IRRDB member countries by following a sequence of activities from data collection, analysis and interpretation of results. This proposal has been submitted for the approval of the financial committee. 2. Publication of a Special Issue in RRISL Bulletin - International Forum on Current Socio-economic Issues and their Impact on Rubber Production Sector Performance 3. Publication of a guidebook on appropriate use of Statistical Methods in Socio-economic research in the rubber sector List of publications on Socio-economics by member countries of IRRDB China: Liu Ruijin, Yang Lin and Wei Hongjie (2017). “Study on the Co-movement Behaviors of Nntural and Synthetic Rubber Price and Exchange Rates” (Chinese). Issues of Forestry Economics, 37(2): 61-70. He Changhui. (2018). Capacity Forecast of Rubber Planting Industry in China. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 30. India: Rubber cultivation driven socio-economic development of tribal communities in Odisha, T. Siju; Binni Chandy and James Jacob, Rubber Science 31 (2), Pages 140-151.

Trends in seasonality of natural rubber production in major producing countries: a disaggregate level analysis, S.Veeraputhran & Shammi Raj and K. Tharian George, Rubber science 30 (1), Pages 66-75.

Incremental economic worth of the first high yielding Indian hybrid clone, RRII 105 over RRIM 600, James Jacob and T. Siju. Rubber Science 30 (3),Pages 193-200.

An analysis of the activities performed by the Rubber Producers’ Societies and strategies to improve their performance, T. Siju; Joby Joseph; Binni Chandy; James Jacob and K.N. Haridas, Rubber Science 31 (1), Pages 60-68.

A road map for attaining self-reliance in natural rubber production in India by 2030. James Jacob; Joby Joseph and T. Siju, Rubber Science 31 (2), Pages 83-91.

Over-dependence of Indian rubber industry on imported natural rubber: the question of long-term sustainability, Joby Joseph and James Jacob, Rubber Science 31 (1), Pages 1-9.

India’s rubber sector under trade pacts: a review of ASEAN India FTA. By Joby Joseph in National Workshop on International Agricultural Trade and Free Trade Agreements: Towards Livelihood Security of Farmers” Organized by State Agricultural Prices Board in association with WTO Cell, Department of Agriculture Development and Farmer’s Welfare, Government of Kerala, held at Trivandrum, Kerala, India. 26 – 27 June 2018.

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Binni Chandi and James Jacob. (2018). Rubber smallholders in India: Socio-economic issues, challenges and opportunities. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 31. Indonesia: Titik Widyasari and Akhmad Rouf (2017). Productivity Effect on the Cost Price of Rubber Estate in Central Java. Jurnal Penelitian Karet, 2017, Vol 35, No 1. Titik widyasari (2017). The Role of Rubber Based Plantations (Hevea brasiliensis Muel Arg) in the Supplying of Horticultural Commodities. National seminar of Sebelas Maret University, Solo, Central Java, Indonesia, March 30, 2017. Dwi Shinta Agustina and Eva Herlinawati (2017). Comparison study of Investment Feasibility of GT 1 and PB 260 Clones at Various Level of Price and Economic Ages. Indonesian Journal of Natural Rubber Research, 35(1). Dwi Shinta Augustina. (2018). Enduring Low Rubber Prices and Socio-economic Problems faced by the Rubber Smallholders in South Sumatra. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 27. Cote d’Ivoire: ATSIN G. J. O., SOUMAHIN E. F., MAHYAO A., OKOMA K. M., KOUAKOU T. H. and OBOUAYEBA S. (2017). Incidence of latex harvesting technologies on agronomic and physiological parameters and profitability of some rapid metabolic class clones of rubber tree (Hevea brasiliensis) in southwestern Côte d’Ivoire, Int. J. Biol. Chem. Sci. 11(5): 2209-2224 Sri Lanka: Munasinghe, E. S. and Rodrigo, V. H. L. (2018). Rubber cultivation shows positive impact on rural livelihood in the Eastern province of Sri Lanka. Dissemination of Research Findings - Private Sector Engagement for Sustainable Development in Sri Lanka: Evidence-Based Policy and Practice. Sri Lanka Institute of Development Administration. 41- 44. Munasinghe, E. S., Rodrigo, V. H. L., Jayathilake, P. M. M and Piyasena, N. M. (2018). Rubber cultivation shows positive impact on rural livelihood in the Eastern province of Sri Lanka. Proceedings of the Second International Agricultural Research Symposium 2018. Sri Lanka Council for Agricultural Research Policy. 126. Munasinghe, E. S. and Rodrigo, V. H. L. (2018). Sri Lankan experience in rubber cultivation in non-traditional areas. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 29. Gunarathne, P.K.K.S., D.M.A.P. Dissanayake, W. Wijesuriya and R.A.D. Ranawaka. (2018). Livelihoods of rubber latex harvesters in Kegalle District: Comparative analysis of harvesting own lands versus operating on hired basis. Proceedings of the Second International Agricultural Research Symposium 2018. Sri Lanka Council for Agricultural Research Policy. 141-142. Dunuwila, P, Rodrigo, V.H.L. and Naohiro Goto (2018). Financial and environmental sustainability in manufacturing crepe rubber in terms of material flow analysis, material flow cost accounting and life cycle assessment. Journal of Cleaner Production, 182, 587-599. Kudaligama, K.V.V.S., Rodrigo, V.H.L., Randunu, R.P.S., Perera, M.K.P. and Madushani, P.D.T.L. (2017). A low frequency harvesting system exclusively for smallholder rubber growers being employed off

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farm. International Rubber Conference 2017, International Rubber Research and Development Board, Grand Sahid Jaya Hotel, Jakarta-Indonesia, 18–22 October 2017. Dissanayake, Anura. (2018). Strategic technology transfer methods: Experience from the smallholder rubber sector of Sri Lanka. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 32. Sankalpa J.K.S., Wijesuriya B.W., Karunaratne S.B. and Ishani P. G. N. (2017). Use of GIS Geographically Weighted Regression to determine productivity and their driving forces: A case study from the Kalutara district of Sri Lanka, Applied Economics and Business 1(2), 12-27, Available from http://aeb.wyb.ac.lk/index.php/v21p2/ Sankalpa J.K.S., Wijesuriya, W., Karunaratne, S. and Ishani, P.G.N. (2017). A comparison of spatial interpolation methods in developing yield maps of Natural Rubber (NR) for precision crop production: A case study from smallholder rubber lands in Kalutara District Sri Lanka, Proceedings of the International Statistics Conference, Sri Lanka. Ishani P.G.N., Sankalpa J.K.S. and Wijesuriya, W. (2018). Sri Lankan rubber exports performance by major destinations using shift-share analysis, 11th Annual Research Forum, Sri Lanka Agricultural Economics Association, Kandy, Sri Lanka. Sankalpa J.K.S., Wijesuriya W., Karunaratne S.B. and Ishani P. G. N. (2017). Use of GIS Geographically Weighted Regression to determine productivity and their driving forces: A case study from the Kalutara district of Sri Lanka (Abstract only), International Rubber Conference, Jakarta, Indonesia, 2017. Malaysia: Siti Hawa Sulong. (2018). Socio-economic problems of the Rubber Smallholders and their solutions: Malaysia’s Experience. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 26. Cambodia: Sreynet Srun. (2018). Current Socio-economic Impacts Caused by Low Prices of Natural Rubber in Cambodia. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 28. Thailand: Preprame Tassanakul. (2018). Introducing RSS GMP Premium Grade for Rubber Farmer Group Sustainability. International Workshop on Statistical Applications in Socioeconomic Research and Forum on Current Socioeconomic Issues and their Impact on Rubber Production Sector Performance. 1-5 October 2018, Rubber Research Institute of Sri Lanka. 33.

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IRRDB ACADEMY

TRAINING FELLOWSHIP

REPORT ON IRRDB TRAINING FELLOWSHIPS 2017 “INCREASING PRODUCTIVITY THROUGH THE ADOPTION OF GOOD AGRICULTURAL PRACTICES”

RUBBER RESEARCH INSTITUTES OF SRI LANKA (RRISL) 08th - 14th OCTOBER 2017

AND

IRRDB/IRRI INTERNATIONAL RUBBER CONFERENCE 2017 “CURRENT AND FUTURE CHALLENGES IN RUBBER INDUSTRY: CAPTURING OPPORTUNITIES IN THE

GREEN ECONOMY” IN CONJUNCTION WITH WORLD PLANTATION CONFERENCES AND EXHIBITION 2017

PLANTATION IN THE CURRENT AND FUTURE CHALLENGES JAKARTA, INDONESIA

15th - 20th OCTOBER 2017

by MUHAMMAD ZAHIDIN BIN RAZMAN

RESEARCH OFFICER (Q44) MRB SOUTHERN REGION

EXTENSION AND DEVELOPMENT DIVISION (BPP)

International Training Fellowship Programme on Sustainable Rubber Cultivation Academy of International Rubber Research and Development Board 1.0 INTRODUCTION The natural rubber industry is going through very challenging times. It is important for the rubber research institutes, all members of the IRRDB, to prepare the young scientists to rise up to the challenge. This is very important considering that natural rubber is mostly produced by smallholders. To address and to implement this issue, the first Academy Fellowship Training Programme held in Rubber Research Institute of Sri Lanka has brought together researchers in various disciplines and plantation sector personnel to share their expertise and of course to transfer of knowledge among each other. This training programme has a wide range of topics related to the challenges faced by the industry to ensure sustainable rubber plantations mostly common to all countries. On the other hand, this training programme and congregation will provide a favorable atmosphere for all researchers to participate in meaningful discussions and interactions to enhance knowledge and understanding of the underlying issues. 2.0 OBJECTIVES

2.1 To share and focus on increasing productivity through adoption of Good Agricultural Practices among participants. 2.2 To study and discuss the knowledge and experience on good agronomic practices, plant protection, latex harvesting technology and intercropping practices to enhance income of rubber smallholders. 2.3 To build network and better communication among young researches in different countries

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3.0 LIST OF PARTICIPANTS

No Name Position Organization Email 1 Dr Erivaldo Jose Scaloppi

Junior Scientific Researcher

Center of Rubber Tree and Agroforestry Systems, Agronomic Institute, Brazil

scaloppijr@yahoo.com.br

2 Mr. R.C. Jan Balzatar Evangelista

Researcher University of Southern Mindanao, Philippine

rcjevangelista@gmail.com

3 Ms. Martini Aji Researcher Sembawa Research Center, Indonesia

martiniaji.ma@gmail.com

4 Mr. Zhang Xicai Assoc. Prof Rubber Research Institute of CATAS, China

hawk771@126.com

5 Mr Muhammad Zahidin bin Razman

Researcher Malaysian Rubber Board, Malaysia

zahidin@lgm.gov.my

6 Dr. Okundia Rex Osabuogbe

Researcher Rubber Research Institute of Nigeria

rexokuns@yahoo.com

7 Mrs. Thamonwan Thonusin

Agricultural Researcher

Buriram Rubber Research Center, Thailand

wittayasrrc@hotmail.com

8 Dr. (Ms.) Nyaka Ngobisa Aurelie epse Mandengue

Regional Chief of Center Research and Innovation

Institute of Agricultural Research for Development (IRAD), Cameroon

boulnyaka@yahoo.com

9 Mrs. P. V. Asiri Anushka Researcher Rubber Research Institute of Sri Lanka

anushpva83@gmail.com

10 Mr. San Prak Researcher Cambodian Rubber Research Institute, Cambodia

prak.sann.ps@gmail.com

11 Dr. Thomson Abraham Researcher Rubber Research Institute of India

thomson@rubberboard.org.in

12 Mr. Pyitt Soann Estate Manager

Mynmar Rubber Planters & Producers Association

pyittsoann@gmail.com

13 Mr. Dang Quoc Thong Deputy Director

Rubber Research Institute of Vietnam

quocthong1980@gmail.com

14 Mr. Jose Alberto Mota Leon

Plantation Supervisor

Company of Gremial de Huleros, Guatemala

jamltico@gmail.com

15 Mr Mauro Francisco Leon Cifuentes

Plantation Supervisor

Company of Gremial de Huleros, Guatemala

mauleci@gamil.com

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4.0 TENTATIVE PROGRAMME DAY O1 MONDAY - 9th OCTOBER 2017 0815 - 0845 Registration of fellowship participants at the auditorium of RRISL INAUGURAL SESSION 0900 - 0915 Welcome of delegates

Mr. N V T A Weragoda, Chairman, Rubber Research Board of Sri Lanka 0915 - 0930 Introduction to the IRRDB Academy

Datuk Dr. Abdul Aziz S A Kadir, Secretary General of IRRDB 0930 - 0940 Introductory Remarks on the World;s First Rubber Research Institute

Mr Dhammika Ranatunga, Acting Director, RRISL 0940 - 0950 Introduction to the fellowship programme

Dr Lakshman Rodrigo, Additional Director, RRISL 0950 - 1030 Presentation of country reports by the participants 1030 - 1100 Tea 1100 - 1215 Presentation of country reports by the participants 1215 - 1245 Guest Speech 1 : Natural Rubber Productivity : Breaking the yield barrier

Dr. Ramli Othman & Dr. Abdul Aziz S A Kadir END OF INAUGURAL SESSION 1215 - 1415 Lunch 1415 - 1515 Guest Speech 2 : Reduce Immaturity Period and increase productivity :

GAP from nursery to field Latex harvesting technology : Secret to saving bark, reducing wastages and extanding economic life Mr. Tajuddin Ismail

1515 - 1530 Tea 1530 Field Visit to sites of germplasm conservation & IRRDB clone exchange

repositories END OF DAY 1

DAY O2 TUESDAY - 10th OCTOBER 2017 TECHNICAL SESSIONS 0830 - 0840 Research highlights in Rubber Agronomy

Dr (Mrs) Proyani Sevaniratne, Deputy Director Research (Biology), RRISL 0840 - 0920 Breeding Rubber for Diverse Needs : What is already Achieved and what

to be achieved Dr (Mrs) Samanthi Withanage, Head, Department of Genetics & Plant Breeding, RRISL

0920 - 0940 Agro-climatic conditions in rubber growing areas of Sri Lanka Dr (Mrs) Wasana Wijesur iya, Principal Research Officer, Biometry Section of RRISL

0940 - 1020 Meeting the present day challenges in planting material production Dr (Mrs) Priyani Senaviratne, Deputy Director Research (Biology), RRISL

1020 - 1100 Latex harvesting for sustainable plantation management Dr Chamil Nayanakantha, Head, Department of Plant Science, RRISL

1100 - 1130 Tea

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TECHNICAL SESSIONS 1130 - 1210 Cost Effective Application of Good Agricultural Practices (GAP) in rubber

lands Dr (Mrs) Rasika Hettiarachchi, Senior Research Officer, Department of Soils and Plant Nutrition, RRISL

1210 - 1250 Biochemistry and Physiology of rubber tree and latex Dr (Mrs) Sagari Kudaligama, Senior Research Officer, Department of Biochemistry and Physiology, RRISL

1250 - 1415 Lunch TECHNICAL SESSIONS 1415 - 1455 Recent Advances in plant protection and way forward

Dr (Mrs) Sarojani Fernando, Principal Research Officer, Department of Plant Pathology and Microbiology, RRISL

1455 - 1530 Guest Speech 3 : History of Natural Rubber in Asia Dr. C. K. Jayasinghe, IRRDB Fellow

1530 Visiting laboratories at Dartonfield and interacting with Scientists of RRISL

1700 Tea END OF DAY 02

DAY O3 WEDNESDAY - 11th OCTOBER 2017 TECHNICAL SESSIONS 0830 - 0900 Multifaceted approach in promoting rubber in non-traditional areas

Dr Lakshman Rodrigo, Additional Director, RRISL 0900 - 0930 Carbon Trading to Value the environmental benefits of rubber

Dr (Mrs) Enoka Munasinghe, Principal Research Officer, Adaptive Research Unit, RRISL

0930 - 1015 Guest Speech 4 : Rubber research, research allocation and return on investment Mr Lakna Paranawithana, Advisor to the Hons. Minister of Plantation Industries and Sneior Advisor, Sri Lanka Society of Rubber Industry

1015 - 1100 Tea 1100 - 1110 Research Highlights in Rubber Technology 1110 - 1150 Cost Effective Approaches for raw rubber processing

Dr Susantha Siriwardena, Deputy Director Research (Technology), RRISL 1150 - 1230 Homemade Rubber Products; value addition at farmer level

Dr (Mrs) Dilhara Edirisinghe, Head, Department of Rubber Technology and Development, RRISL

1230 - 1400 Lunch 1400 - 1440 Effective Technology Transfer process to achieve high productivity in the

smallolder sector Dr Anura Dissanayake, Head, Advisory Services Department, RRISL

1440 - 1530 Guest Speech 5 : Next Generation Technology Transfer Dr Jacob Mathew, IRRDB Fellow

1530 - 1600 Tea

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1930 Dinner hosted by the Rubber Research Institute of Sri Lanka

END OF DAY 03

DAY O4 THURSDAY - 12th OCTOBER 2017 FIELD VISITS 0730 Visit to Sapumalkanda estate, Lalan Rubbers, Deraniyagala to

experience cost effective plantation management Lunch at Sapumalkanda estate

1500 Visit to a glove factory at Avissawella Stay in Dambulla

END OF DAY 04

DAY O5 FRIDAY - 13th OCTOBER 2017

FIELD VISITS AND SIGHT SEEING Visit to Sigiriya, the Lion Rock Kingdom , Visit to Cave Temple, Dambulla Visit to Temple of Sacred Tooth Relic, Kandy Stay in Kandy END OF DAY 05

DAY O6 SATURDAY - 14th OCTOBER 2017

FIELD VISITS AND SIGHT SEEING Visit to Sanquhar estate, Gampola , Lunch at Royal Botanical Garden, Peradeniya Visit to Smallholding and a processing center Closing remarks Stay in Colombo END OF DAY 06

DAY 07 SUNDAY - 15th OCTOBER 2017

TRAVELLING DAY Bandaranaike Airport (Sri Lanka) to Soekarno-Hatta Airport (Indonesia) Stay in Bogor END OF DAY 07

DAY 08 MONDAY - 16th OCTOBER 2017 0900 - 1100 Visit to Indonesian Rubber Research Institute, Bogor 1100 - 1300 Visit to Pt. Supervulkanin Adijaya 1300 - 1400 Lunch 1400 - 1700 Visit to Indonesian Rubber Research Institute, Bogor (Laboratories)

Visit to Royal Botanical Garden, Bogor Stay in Bogor END OF DAY 08

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DAY 09 TUESDAY - 17th OCTOBER 2017 0800 - 1300 Visit to Puncak 1300 - 1400 Lunch 1400 - 1800 Departure to Jakarta

Stay in Jakarta END OF DAY 09

DAY 10 WEDNESDAY - 18th OCTOBER 2017 OPENING SESSIONS OF WPLACE 2017 0800 - 0930 Registration 0930 - 1000 Opening Ceremony 1000 - 1030 Coffee/Tea Break 1030 - 1100 Keynote Speech 1 - Coordinating Minister of the Economic Affairs 1100 - 1200 General Topic 1 - Minister of Finance of the RI 1200 - 1310 Lunch Break 1310 - 1320 Welcome address - Dr Karyudi 1320 - 1330 Opening Remarks - Datuk Dr Abdul Aziz S A Kadir 1330 - 1350 The Role of the International Tripartite Rubber Council (ITRC) in

Stabilizing NR Prices Dr. Chairil Anwar

1350 - 1410 Demand and Supply of Natural Rubber Salvatore Pinizzotto

1410 - 1430 An Insight to Rubber Price Movements from Macro Financials Dar Wong

1430 - 1450 Policy Options for Managing Farm Income under Market Uncertainties Dr. Nguyen Ngoc Bich

1450 - 1510 Natural Rubber : Assesing its Potential as a Sustainable Crop Moenardji Soedargo

1510 - 1525 Discussion 1525 - 1545 Coffee/Tea Break 1545 - 1605 Recent Developments and Growth Trends in The Automobile and Tyre

Industry in India - Implication for the NR Sector Rajiv Budhraja

1605 - 1625 The Rise of the Green Economy : Implications on the NR Industry Prof Dato' Dr. Ahmad bin Ibrahim

1625 - 1645 Current Trends in Latex Dipped Goods Industry Prof Dr. Ho Che Cheong

1645 - 1705 Manufacturing High Quality Engineering Rubber Product for Protection of Structures and Buildings Or Tan Teng

1705 - 1725 Natural Rubber in Engineering Application Dato' Dr. Kamarudin Ab Malek

1725 - 1745 Enabling Precision Farming with Drone Technology for Local Crops Arif Makhdzir

1745 - 1800 Discussion

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DAY 11 THURSDAY - 19th OCTOBER 2017 0815 - 0825 Developmental Dynamic of Activity and Expression of Sucrose-

Metabolizing Enzymes in Hevea brasiliensis Leaves Tang Chaorong

0825 - 0835 Soil under Hevea in Peninsular Malaysia : Identification and Distribution of Clay Minerals Wan Mohamed Noordin Wan Daud

0835 - 0845 Nitric Oxide Improves Growth and Physiological Attributes of Rubber (Hevea brasiliensis) under Drought and Heat Stress Conditions Chamil Nayanakantha

0845 - 0855 Use of GIS Geographically Weighted Regression to Determine Natural Rubber Productivity and Their Driving Force : A Case Study from the Kalutara District of Sri Lanka Sajeep Sankalpa

0855 - 0905 Improving Soil Chemical Properties and Growth Performance of Hevea brasiliensis through Basalt Application Mohd Shafar Jefri

0905 - 0915 Tracking Carbon from Photosynthesis to Latex with 13C Field Labelling Experiment Philippe Thaler

0915 - 1000 Discussion 1000 - 1030 Coffee Break 1030 - 1040 Molecular Cloning and Characterization of S-Adenosylmethionine

Decarboxylase Gene in Rubber Tree (Hevea brasiliensis) Li Dejun

1040 - 1050 Molecular Evolution, Cloning and Characterization of SnRK2 Gene and its Expression Profiling in Rubber Tree Dong Zhang

1050 - 1100 Genome-wide Identification of Cis-elements in Promoter Region of Hevea brasiliensis Genome Xi Huang

1100 - 1110 Analysis of Molecular Markers Linked to Corynespora Leaf Fall Disease Resistance in Rubber Plants Fetrina Oktavia

1110 - 1120 Protoplast Fusion Between PR 107 and Reyan 8-79 by using PEG Dai Xuemei

1120 - 1130 Characteristic, Physiology and Yield Potential of RRIT 251 and RRIT 408 Clone Ratchanee Ratanawong

1130 - 1200 Discussion 1200 - 1300 Lunch Break 1300 - 1310 An Exploratory and the Development of Rubber Spectral Library for Different

Rubber Clone Mohd Hafiz Mohd Hazir 1310 - 1320 Analysis of Phenotypic Diversity of Selected Hevea accesions from IRRDB

1981 Germplasm Collection Conserved in Sri Langka PVA Anushka

1320 - 1330 The Potential Genotypes with High Timber Volume from Six Species in the 1995 RRIM Hevea Germplasm Collection Mohd Adi Faiz Ahmad Fauzi

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1330 - 1340 Study on Wood Anatomy and Mechanical Properties of Clone RRIM 3001 with Protuberances Mohd Syolahuddin Mokhter

1340 - 1350 The Performance of IRR Series Rubber Clones on Trial Scale and in Commercial Plantations Sekar Woelan

1350 - 1415 Discussion 1415 - 1425 Opportunities and Challenges in Green Plant Protection Development of

Natural Rubber in China Huang Guixiu

1425 - 1435 Functional Analysis of HbMlo12 in Powdery Mildew Infection and Stress Response in Rubber Tree Hevea brasiliensis Muell Arg Yu Zhang

1435 - 1445 Evaluation of Phylloplane, Cauloplane and Rhizosphere Bacteria for Multiple Antagonism Against Five Major Pathogens of Rubber (Hevea brasiliensis) C Kuruvilla Jacob

1445 - 1455 Characterization of Pestalotiopsis microspora, Causal Agent of Leaf Blight on Rubber (Hevea brasiliensis) in Cameroon Nyaka Ngobisa

1455 - 1515 Discussion 1515 - 1545 Coffee Break 1545 - 1555 A Study on Early Cold Responsive Signaling Pathways in Rubber Tree

Cheng Han 1555 - 1605 Distribution of Glyphosate-Resistance Goosegrass (Eleusine indica Edison Purba 1605 - 1615 The Innate Immunity Responses in Rubber Tree

Hongli Luo 1615 - 1625 Biological Control of Corynespora cassiicola Causing Corynespora Leaf Fall

Disease (CLF) on Rubber Tree by Bacillus thuringiensis (T3) Pham Thi Ngoc Giau

1625 - 1635 Pathogenicity Test of Lasiodiplodia theobromae Isolates from Six Host Plants and Their Phylogeny Analysis Tri Rapani Febbiyanti

1635 - 1655 Discussion 1655 - 1800 Closing

DAY 12 FRIDAY - 20th OCTOBER 2017

0830 - 0900 Keynote Speech 2 Minister of Industry of the RI

0900 - 1015 Session 1 : Downstream Industry Development

1015 - 1130 Session 2 : Upstream Industry Development

1130 - 1300 Lunch Break and Exhibition Visit 1300 - 1415 Session 3 : Plantation Under Climate

Change 1415 - 1600 Session 4 : Nature Resource Genetic

USA 1600 - 1630 Closing Ceremony END OF PROGRAMME

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Training Fellowships IRRDB 2017 in Sri Lanka On 8th October 2017, my journey has begun from Kuala Lumpur International Airport 2 (KLIA2) to Bandaranaike Airport, Sri Lanka and it took about three and half hours flight. From Malaysia, there were three of us, Datuk Dr Abdul Aziz S A Kadir, Mr Haji Tajuddin Ismail and myself. In Sri Lanka, we were welcomed and entertained by Dr Lakshman Rodrigo and Mr Kalum. We have been brought to the Rubber Research Institute of Sri Lanka (RRISL) by van and stayed at Laya Leisure Hotel, which is located at Kukuleganga, about 15 km away from RRISL. The Training Fellowship Programme started with welcome speech by Mr. N V T A Weragoda, Chairman of Rubber Research Board of Sri Lanka, Introduction to the IRRDB Academy by Datuk Dr Abdul Aziz S A Kadir, Introductory Remarks on the World’s First Rubber Research Institute by Mr Dhammika Ranatunga, Acting Director of RRISL, Introduction to the fellowship programme by Dr Lakshman Rodrigo, Additional Director of RRISL and presentation of country reports by all participants. There were 15 participants from 14 countries were selected in this programme. The first session started with lecture by Mr Haji Tajuddin Ismail and he talked about reduce immaturity period and increase productivity and also latex harvesting technology. In his talk, he summarized that all trees should be of the right stage before they can be opened for tapping, all tappers should undergo formal training and only qualified tappers should be allowed to tap rubber trees, low frequency tapping d/4 is highly recommended, latex DRC must always be maintained over 30% and latex stimulants should be used correctly and judiciously and of course, smallholders should be well exposed to appropriate latex harvesting technologies. In the evening, we have been visited to the sites of germplasm conservation and IRRDB clone exchange repositories. The germplasm collection was brought to Sri Lanka in mid ‘80s and established in the Neuchatel estate, Dickhena Division belongs to Estate Plantation Corporation and now it is more than 30 years old. Sri Lanka has already received about 10,000 genotypes and it occupies nearly 13.5 ha of land. This repository is located in the Katutura District in the Western Province and its approximately 38 km distance from RRISL, Dartonfield. In this repository, vigorous clones have ever grown to the extent of hindering the less vigorous ones. This leads to loss of some of these less vigorous genotypes in the collection and continuously paying attention to fill these vacancies. Having all the genotypes in one location is also a risk. Hence, the duplication of collection in other locations is of vital importance RRISL has already planned to duplicate the germplasm population at least in two more locations, preferably one in the wet area and the other in a dry area with normal density used in commercial plantations. This will enable to do future evaluations for economically important traits as well as the maintenance of material without any losses. On 10th October 2017, there were nine lectures presented by various expertise of researchers from Sri Lanka and the first lecture was delivered by Dr (Mrs) Priyani Se. She talked about the research highlights of Agronomy Departments. There are 10 departments namely Genetics and Plant Breeding, Plant Science, Soils and Plant Nutrition, Plant Pathology and Microbiology, Biochemistry and Physiology, Advisory Services, Adaptive Research Unit, Biometry Unit, Agricultural Economics Unit and Audio Visual Unit. In the history aspect of breeding, Sir Henry Wickham (Naturalist) collected around 70,000 seeds from region of upper Amazon in Brazil. Then about 2700 seeds were germinated in Kew Botanical Garden in UK and about 1919 seedlings were brought to Sri Lanka (Figure 1-6).

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Figure 1: Development of Breeding Objectives with diverse needs

Figure 2: The RRISL Clone Recommendation 2013

Figure 3: Yield (gtt) of RRIC 102 Figure 4: Yield (gtt) of RRIC 121

Figure 5: Average Yield (gtt) of RRIS 203 Figure 6: Average Yield (gtt) of RRIS 208 In Sri Lanka, they also studied about Agro climatology where the main aim in this study is to apply climatological information for the purpose of improving farming practices and increasing agricultural productivity both in terms of quantity and quality. The climatic zones of Sri Lanka are

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divided into two categories namely based on rainfall and based on elevation. In term of rainfall, there are 3 categories, dry zone (< 1750mm), Intermediate Zone (1750 - 2500mm) and Wet Zone (> 2500mm) whereas elevation, divided into three categories, low country (< 300m), Mid Country (300 - 900 m) and Up Country (> 900m). There are 46 regions; 15 regions in Wet Zone, 20 regions in Intermediate Zone and 11 regions in Dry Zone.

Figure 7: The Climatic Zones in Sri Lanka In the aspect of planting material production, RRISL made a study in order to improve further in planting materials and they did rejuvenation mature plants. There are various morphological characteristics to detect Juvenility, namely colour, angle of the shoot and inter-nodal length. The Juvenile planting materials showed fast growth rate, higher girth and high yield. In term of quality control, all rubber nurseries are inspected by RRISL for quality assurance of plants. There are four inspection rounds and four reports are done by RRISL the final report is required to claim for subsidy to make sure that they have used good quality plants. The RRISL also touched about the topic of cost affective application of Good Agricultural Practices (GAP) in rubber lands. They did bio fertilizer, cover crop management, bio char application, organic manure, slow release fertilizer and the combine use of agro management practices. The bio film bio fertilizer is less bulky and easy to handle, low cost about Rs 0.50/plant (RM 0.01/plant) and enviromenta friendly. The advantages are N² fixation, biosolubilization of rock phosphate and production of plant growth promoting hormones. The results of using the biofilm biofertilizers are improving soil nutrient availability, decreasing bulk density, improving microbial biomass carbon and enhance aggregate stability. Biochar is carbon rich material, it is produced from the slow pyrolysis of biomass, highly porous, usually alkaline, high specific surface urea and varied with the source material and condition provided by the pyrolysis process. The RRISL also introduced the fertilizer encapsulated coir blocks for immature Hevea at field conditions. Only one application per year and it reduce labour cost associated with fertilizer application by 75%, reduce fertilizer cost by 20%, minimize fertilizer wastage as environmentally pollution, maximize fertilizer use efficiency and valuable product from cheap industrial waste mineral (Figure 8).

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Figure 8: Fertilizer Encapsulated coir blocks for Immature Hevea In the aspect of Carbon Trading to value the environmental benefits to rubber presented by Dr Enoka, she mentioned about the climate and climate change. The climate is a measure of the average pattern of variation in temperature, humidity, atmospheric pressure, wind, precipitation, atmospheric particle count and other meteorological parameters in a given region over long periods of time. The climate change is a significant and lasting change in the statistical distribution of weather patterns over periods ranging from decades to millions of years. There are two factors that determine the climate change, namely natural and human origin (Figure 9).

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Figure 9: Factors determine Climate Change (Natural Origin) The objectives of international interventions such as United Nations Frame-Work Convention on Climate Change are to stabilize GHG concentration in the atmosphere, to prevent anthropogenic interference with the climate system and to allow for ecosystems to adapt naturally to climate change, ensure food production is not threatened, enables economic development to proceed in a sustainable manner. The next topic that presented and discussed in this training programme was very important part of the rubber industry, namely Next Generation Technology Transfer, presented by Dr Jacob Mathew, Fellow of IRRDB. The definition of TOT is anyr information or technology as well as innovation that come out from researchers, should be transferred to secondary user, known as farmers or rubber industry through extension system. Nowadays, the world has become modern and futuristic in term of communication and understanding, and these modernization should be a part of our methodology of technology transferred to the smallholders. The existing extension delivery system or known as conventional methods have to be changed to the alternative and modern system. The transfer of technology process should be changed from conventional method to alternative methods (Figure 14). Nowadays, we still use conventional methods like individual or group contact, mass contact and communications by bulletin information, newsletter and articles rather than alternative methods such as websites, CD ROM, knowledge portals, interactive webs, video conference and mobile apps (Figure 15). These alternative methods will change the existing smallholders’ thinking and will attract more youngsters to be actively participate and join the rubber industry.

Figure 14: The Cone of Learning in Conventional Methods

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Figure 15: Alternative Methods by using Mobile Apps/Website In a part of technology transfer programme , RRISL also did the multifaceted approach in promoting rubber in non-tradisional areas. The impacts are come from different points of view, such as government, it will aspect more natural rubber can be exploited to product manufacturers, more revenue to government and it will contribute the sustainability of economy. From the point of view of researchers, it will give tangible deliverables whereas from grower, they will say new income avenue with stability and livelihood improvement. There are a lot of approaches that should be done in expansion process, such as be vigilant on risk factor (disease), development of agronomic protocols in participatory approach, socio economic assessments, social interactions with farmer community, institutional support for mutual benefits and public awareness. The non-traditional areas in Sri Lanka is totally differences from traditional areas like different in climate, very remote and less facilities, the farmers have no knowledge on rubber, different priorities at farmer level, lack of institutional support and politic will and very limited funds. In the field activities, the first step is a preliminary work such as interviews with key informants, site visits and collaboration from relevant parties. The next step is on the aspect of agronomy trials on farmer’s field like fertilizer and other GAP and come out with agronomic assessments. The same thing goes to sociological assessments by doing SWAT and Livelihood Analysis. In term of transfer of technology, RRISL did special programmes to get the farmer’s acceptance especially in this non-traditional areas like interaction in on-farm activities, participation in social events, awareness programmes on different aspects and also give training to the smallholders (Figure 16).

Figure 16: Programmes that involve rubber smallholders in non-traditional areas

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On the day 4, we managed to visit Sapumalkanda Estate, which is located in Deraniyagala, in the Sabaragamuwa Province and it belongs to Lalan Group of Plantations. Lalan group is a diversified conglomerate with expertise in over 12 sectors and has been operations since 1940 with branch offices in UK and USA. Lalan has an extensive global network encompassing pan-industry interests in rubber, tea and other plantation crops (such as cardamom, cinnamon, coffee and stevia), reforestary, renewable energy, manufacturing and service industries, engineering and the leisure sector. Established three generations ago in Sri Lanka, the group which started primarily as a rubber commodity trading operation, has expanded, developed and modernized its operational modalities in keeping with global changes. The vision of this company is to become a world class natural rubber plantations management company by 2014. One of the strategies is to preserve, nurture and protect the environment while harvesting the forestlands with productive plants that are inherent to fertile and rich soil of Sri Lanka. The total area is about 6900 hectares and manages over 13 estates include the Sapumalkanda Estate which is about 2000 hectares. The production of latex is over 4.5 million kg. Lalan’s experience in manufacturing crepe rubber extends over half a century. Putting that expertise into practice, the company refurbished and modernized an existing rubber factory at the Sapumalkanda Group. The latex crepe production is about 75,000 kg/month and 80,000 kg/month for RSS Rubber. Lalan’s centrifuging operations are a crucial link in the supply chain, providing centrifuged latex for the production of gloves and latex mattresses. The two centrifuging plants located in the central rubber growing areas of the country work around the clock and have a capacity to centrifuge 10,000 liters of field latex a day. There are about 750,000 kg dry per month of centrifuge latex production. Centrifuged latex produced in Lalan’s factories is exported to Malaysia, New Zealand, Egypt and Belgium. Besides that, the company also produces and exports nearly 1 billion disposable examination gloves, 90 million household gloves and 15 million industrial gloves in 6 factories (Figure 10a - f).

Figure 10a: Nursery Figure 10b: Top Management

Figure 10c: Bush Nursery Figure 10d: Rubber Plantation

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Figure 10e: DRC Determination Testing

Figure 10f: Latex Crepe We were also managed to visit Sanquhar Estate, rubber plantations in high elevations which is located in Gampola District. With the move of the rubber plantation to non-traditional areas, the researchers of RRISL have considered both high elevations area and comparatively drier regions of the country, with the objective of expanding the rubber cultivation to other areas as a solution for the declining extents of rubber plantation in the country, especially in the wet zone. This decline in extent cannot be ruled out in the traditional areas due to urbanization, industrialization and crop diversification. With this objective, an experiment was established in 1996 with 4 clones; namely RRIC 100, RRIC 102 and RRIC 130 which are tolerant to Oidium hevea with a less tolerant clone, PB 260. This trial was established in the Sanquhar Estate (7’ 25’N & 80’ 60’ E) where the elevation ranges from 600 - 900 m above mean sea level. This area belongs to the Mid Country Intermediate Zone with an annual rainfall ranging from 2000 - 2500 mm. Based on the performance, viz. growth, tolerance to leaf diseases and dry rubber yields the clones, RRIC 100, RRIC 102 and RRIC 103 have performed better than the clone PB 260, and these 3 clones have been recommended for planting in areas tested in this study (Figure 11).

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Figure 11 The last two days before leaving Sri Lanka, we were brought to visit to some historical places of Sri Lanka. One of places that we have visited was Kandy the Temple of Sacred Tooth Relic. Sri Dalada Maligawa or the Temple of the Sacred Tooth Relic is a a Buddhist temple in the city of Kandy, which houses the relic of the tooth of the Buddha. Dambulla cave Temple also known as the Golden Temple of Dambulla is a World Heritage Site in Sri Lanka. It is situated in the Central part of the country. This site is located about 148 km east of Colombo and 72 km north of Kandy. Sirigiya is an ancient rock fortress located in the Central Province, Sri Lanka. The name refers to a site of historical and archeological significance that is dominated by a massive column of rock nearly 200 m high. King Kashyapa built his place on the top of this rock and decorated its sides with colourful frescoes. He built a gateway in the form of an enormous lion and the name of this place is derived from this structure of the Lion Rock. Sirigiya nowadays is a UNESCO listed World Heritage Site. It is the one of the best preserved examples of ancient urban planning. Paradeniya Royal Botanical Gardens, the finest of its kind in Asia, the largest of the Botanical Gardens of Sri Lanka, couldn’t be better located. It is located at an elevation of 500 m above sea-level, was tightly bounded on three sides by a loop of River Mahaweli (Great Sandy River), the largest river of Sri Lanka (Figure 12).

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Figure 12: Visit to Historical Places in Sri Lanka International Rubber Conference (IRC) 2017 in conjunction with World Plantation Conferences and Exhibition (WPLACE) 2017, Grand Sahid Jaya Hotel, Jakarta, Indonesia On 15th of October 2017, we travelled to Jakarta from Sri Lanka by Sri Lankan Flight which took about 3.5 hours flight. Since this was my first time in Indonesia, I felt very happy and surprised that there are quite similar with Malaysia, especially in their language, culture, food and also Jakarta city as well. After we arrived at the Soekarno-Hatta Airport in Jakarta, we were brought to Bogor, which is known as the rainfall city in Indonesia because of the raining season throughout the year in Bogor. In Bogor, we managed to visit Indonesian Rubber Research Institute (IRRI) and welcomed by the Director of IRRI, Dr Karyudi and briefed by Deputy Director of IRRI, Dr Thomas. Dr Thomas briefed us about the background of IRRI, their researches and their achievements. In human resources, there are four IRRI stations in Indonesia and about 72 researchers in various expertise. There are four recommended clones in IRR Series 100 and 200, namely IRR 104, IRR 112, IRR 208 and IRR 220. The productivity is range from 1.5 to 2.0 tons/ha/year and wood volume is about 0.27 - 0.46 m3/tree. There are 2 clones have been patented, IRR 220 and IRR 230. IRRI also has introduced the PUKALET (Slow Release Fertilizer) in order to enhance efficiency and decrease in interval fertilizer application and it formulized in the form of compound fertilizer with the nutrition composition depending on soil and leaf analysis. In term of disease especially white root disease (WRD), IRRI introduced the Triko SP, which is bio-fungicide to control WRD and it benefit to prevent and cure the disease via control WRD at 1st and 2nd stage. IRRI also did horticulture crops as rubber intercrops such as Curly Chili, Small Chili, Long Bean, Water melon and banana + pineapple in order to fully utilize of land usage and also to enhance smallholder’s income. Recommended tapping system is based on clonal typology which is physiological response to wounded and stimulant. There are 2 type of clones namely quick starter and slow starter. There is also another technology has been introduced by IRRI in term of tapping, which is called as Tapping Quality Controller (Tap-SP). This tools can be used to measure tapping depth, bark thickness, bark consumption, timber wound and tapping angle. IRRI also has introduced BI-CUT and it is specially designed to control bark consumption and tapping depth. In the downstream sector, IRRI has introduced DEORUB as Latex Coagulant. This material produced by pyrolysis process of oil palm shell and it is environmentally friendly. It is an odorless raw rubber material and better quality which is lower production cost about 20% for crumb rubber and it helps RSS production by reducing the drying process from 5 days to 2 days. In term of rubber products, IRRI has introduced Sebutret which is made from coconut fibre plus latex compound. It has been used in mattress and vases. Rubber compound also has been used for reducing soil erosion which is grass seed is mixed with rubber compound and sprayed into soil. This study is collaborated with Road and Bridge Research Center. Besides that, rubberized road also being a major part of collaboration, which is rubberized road, is made from asphalt 3-5%

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additive (rubber). IRRI also give services to the public especially in consultation (feasibility study, tapping system and immature period technology) (Figure 13).

Figure 13: Activities in Indonesian Rubber Research Institute (IRRI), Bogor, Indonesia In the afternoon, we were very lucky to get visited one of the biggest company of retread tyre belongs to PT Supervulkanin Adiajaya, which is located in Bogor, Indonesia. At the initiative (late) Johnson Lumban Tobing, right in 1973 stands the company (factory) tire retreading which later became known as PT Supervulkanin Adijaya. The hope is not grandiose, he just wants to help the owners of modes of transportation in the efficient use of new tires without the slightest neglect of aspects of safety and comfort. The term "retread tire" is the process of attaching a new tire (recondition). The factory located at Jl. Raya Bogor Km. 52 Kedung Halang Bogor employs approximately 200 employees: consisting of 180 male employees while 20 female employees focus on administration. The long journey that we went through and the tidal (political ups and downs) of politics in this country encouraged us to find the right formula for the sustainability of this company. It is true, not always age to be a benchmark of company quality. But our presence for more than 40 years is evidence that ultimately leads us to the point of awareness that the quality of the products produced not only rests on the facilities and raw materials used but also how the quality of its Human Resources. Nevertheless, we recognize that the era of reformism has not fully sided with service industries like ours. Major communities and power elites still discriminate against the existence of vulcanized factories. Logical fallacy terminology and retreaded products are

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considered not to provide safety guarantees for users. This kind of judgment arises because of the lack of access to information about what and how to retread itself. As has been explained that vulcanization in the context of Supervulkanin Adijaya, committed to strive for maximum quality standards, so that the user vulcanization services in the company get three benefits at once: Efficiency, Comfort and Safety. The embodiment of these three aspects, unraveled in the following apects. First, from the use of raw materials. To produce a quality product, we use a consistent and consistent tread compound and gum compound, under the MBS brand; using the weight of the rubber in accordance with the packaging of the casing; use of tread compound and modern technology. Second, supported by modern retreading technology to produce quality products and satisfy consumers. To produce high quality retreads, PT Supervulkanin Adijaya is supported by high tech machinery. Thirdly, our company procedurally also implements the working principles set forth in the OHS (Health and Safety) items - in collaboration with the Bogor Institute Safety strives for a good and proper working behavior. The basic principle of Occupational Health and Safety is not only a guarantee of physical protection of employee activities within the company but more than that is a form of corporate appreciation of the interests and rights to the protection of a comfortable working life. With the creation of a comfortable and safe working atmosphere, employees will also contribute greatly in improving the quality of production, moreover they get support from family at home. Fourth, the most substantive thing we do is concern on Human Resource development. Since 2012 Robinson as the second generation who leads this company trying to how the human resources is not only skilled from technical skills (hard skills) but also make the character as the main base in all production activities in Supervulkanin Adijaya (soft skill). In the context of hard skills, the company engages employees in various trainings and workshops that are relevant to the needs of our company. In addition, improving the quality of human resources is not only on the development of technical skills but also the personality (karekter) -soft skill-character-based training content is packed philosophically. Employees are invited to discuss with the struggle and existence of employees as human beings (family, work) from different angles. This is so that employees have a broad understanding horizon and ultimately they understand the true essence of work. Balancing between hard skills and soft skills is not easy, the principle clash between employee and employee is inevitable. But with Robinson's consistency, continuity and creativity, it gradually shines in Supervulkanin Adijaya-the light is called 'collective conscience' -Durkheim describes the collective consciousness as a system of inter-individual relationships. For Robinson, a combination of human resource development that includes hard skills and soft skills that are the company's unique characteristics (Value proposition) - Building a Humanized Business-That is, the products we offer are not just the result of mere technical skills but are born of a strong personality employees. Robinson's efforts in maintaining the originality of his idea and his expectation of educating employees, was born Vulkanin Learning Institute or Learning Center. Future Learning Center is not only as a womb that gave birth to the awareness of employees of this company alone but also will invite all the relations together experience a philosophical encounter in order to welcome globalization. This International Rubber Conference 2017 was held to share research experience, knowledge dissemination and introduce new technology among researchers, academicians, experts, smallholders, and industrial practitioners involved in the rubber industry. This conference gathered participants from many rubber producing countries to share new findings, latest research work and improvement of technologies. This conference an opportunity to deliberate on

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the current issues and future challenges focusing on the theme “Current and Future Challenges in Natural Rubber Industry: Capturing Opportunities in The Green Economy”. The International Rubber Conference 2017 (IRC) was divided into eight topics and spanning three days (18th – 20th October 2017) in conjunction with World Plantation Conferences and Exhibition 2017 (WPLACE-2017). Overall, there are eight broad topics were discussed includes current supply and demand of world natural rubber, development of downstream rubber industry, agronomy, plant breeding, plant protection, latex harvesting, socio economy and processing technology. Each session consisted of several speakers and was moderated by a chairman. The opening WPLACE-2017 on 18th October 2017 ceremony was inaugurated by His Excellency Dr. Muhammad Jusuf Kalla, Vice President of the Republic of Indonesia (RI) followed by keynote speech, Dr. Ir. Musdhalifah Machmud, Deputy Minister of Food and Agriculture, the Coordinating Ministry of Economic Affairs Republic of Indonesia. The WPLACE-2017 welcoming speech was done by Dr. Ir. Teguh Wahyudi, President Director of PT-RPN. Next speaker was Dr. Mardiasmo, Vice Minister of Finance on general topic of agriculture current status in Indonesia. Then, participants were divided into each crops session of rubber, oil palm, tea, sugar cane, coffee, cocoa and biotechnology. The separate session of International Rubber Conference 2017 (IRC) was held at Puri Ratna Hall and began of welcoming address by Dr. Karyudi, Director of Indonesian Rubber Research Institute. After that, opening remark was given by Datuk Dr. Abdul Aziz SA Kadir, Secretary General of IRRDB. There were two session on the first day with no parallel session. 1. Session 1: Current Supply & Demand of World Natural Rubber The first speaker was Dr. Chairil Anwar as a CEO of IRCO Thailand on the role of the International Tripartite Rubber Council in stabilizing rubber price. The second speaker was Salvatore Pinizzotto, Secretary General of IRSG Singapore, presented supply and demand of Natural Rubber. The third speaker was Dar Wong from Dektos Investment Singapore talked on an insight to rubber price movements from macro financials. The last speaker was Moenardji Soedargo, Chaorman of GAPKINDO Indonesia on natural rubber: assessing its potential as a sustainable crop. 2. Session 2: Development of Downstream Rubber Industry The first speaker was Rajiv Budhraja, Director of ATMA India presented on recent developments and growth trends in the automobile and tyre industry in India – implication for the NR sector. The rise of the green economy: implications on the NR industry talked was given by Prof Dato’ Ahmad Ibrahim, Academy of Science Malaysia as a second presenter. The third speaker was Prof Dr. Ho Che Cheong, Vice Chancellor of First City Univ College Malaysia on current trends in latex dipped good industry. The fourth speaker was Or Tan Teng, CEO of Doshin Rubber Products (M) Sdn. Bhd. Malaysia presented on manufacturing high quality engineering rubber product for protection of structures and buildings. The fifth speaker was Dato’ Dr. Kamarudin Ab Malek (CEO of Tun Abdul Razak Research Center, UK) on Natural Rubber in Engineering Application. The last speaker was Arif Makhdzir, Managing Director of Brain Technologies Sdn. Bhd. from Malaysia on application of artificial intelligance & drone technology in Malaysian plantation.

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On the second day, there were two parallel session and I’m decided to stay at Parallel 1 as it was same hall for my presentation session.

1. Session 1: Agronomy

The first speaker was Tang Chaorong from China on development dynamic of activity and expression of sucrose-metabolizing enzymes in Hevea Brasiliensis leaves. The second speaker was Chamil Nayanakantha from Sri Lanka on Nitic Oxide improves groeth and physiological attributes of rubber under drought and heat stress conditions. The third speaker was Sajeep Sankalpa from Sri Lanka on use of GIS geographically weighted regression to determine nautral rubber productivity and their driving force: a case study from the Kalutara District of Sri Lanka. The fourth speaker was Philippe Thaler from CIRAD, France on tracking carbon from photosynthesis to latex with 13C field labelling experiment. The last speaker was Guilherme Costa from Brazil on natural rubber an opportunity in Brazil.

2. Session 2: Plant Breeding 1 The first speaker was Li Dejun from China on Molecular Cloning and Characterization of S-Adenosylmethionine Decarboxylase Gene in Rubber Tree. The second speaker was Dong Zhang from China. The objectives of their research were to identify and characterize of the sucrose non-fermenting 1-related protein kinase 2 (SnRK2) gene family members in Hevea Brasiliensis. The third speaker was Xi Huang from China on genome-wide identification of Cis-element in promoter region of Hevea Brasiliensis genome. The fourth speaker was Ferina Oktavia from Indonesia on analysis of molecular markers linked to corynespora leaf fall disease resistance in rubber plants. The fifth speaker was Dai Xuemei from China on asymmetric protoplast fusion between PR 107 and reyan 8-79 by using PEG. The last speaker was Ratchanee Ratanawong from Thailand on characteristic, physiology and yield potential of RRIT 251 and RRIT 408 clone.

3. Session 3: Plant Breeding 2

The first speaker was I, Mohd Hafiz Mohd Hazir from Malaysia on an exploratory and the development of rubber spectral library for different rubber clone. The second speaker was PVA Anushka from Sri Lanka on analysis of phenotypic diversity of selected Hevea accessions from IRRDB 1981 germlasm collection conserved in Sri Lanka. The third speaker was Mohd Adi Faiz Ahmad Fauzi from Malaysia on the potential genotypes with high timber volume from six species in the 1995 RRIM Hevea germplasm collection. The forth speaker was Mohd Syolahuddin Mokhter from Malaysia on wood anatomy and mechanical properties of clone RRIM 3001 with protuberance. The last speaker was Sekar Woelan from Indonesia on the performance of IRR series rubber clone on trial scale and commercial plantation.

4. Session 4: Plant Protection 1 The first speaker for this session was Huang Guixiu from China on opportunities and challenges in green plant protection development of natural rubber in China. The second speaker was Yu Zhang representative from China which gave talked on Functional analysis in powdery mildew infection and stress response in rubber tree Hevea brasiliensis Muell Arg. The third speaker was C Kuruvilla Jacob from India on the topic of evaluation of phylloplane, cauloplane and rhizosphere bacteria for multiple antagonism against five major pathogens of rubber. Nyaka Ngobia from Cameroon was the last speaker for this session which present on characterization of pestalotiopsis microspore, causal agent of leaf blight on rubber in Cameroon.

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5. Session 5: Plant Protection 2

The first speaker was Cheng Han from China presented on comparative transcriptome analysis reveals early gene expression profile that contributes to cold resistance in Hevea brasisliensis. Second presenter was Edison Purba from Indonesia on distribution of glyphosate- resistance goosegrass (Eleusine indica) populations and an alternative herbicide (Indaziflam) to control them. The third speaker was Hongli Luo from China on the innate immunity responses in rubber tree. The forth speaker was Pham Thi Ngoc Giau from Vietnam presented on biological control of corynespora cassiicola causing corynespora leaf fall disease (CLF) on rubber tree by Bacillus thuringiensis (T3). The last speaker was Tri Rapani Febbiyanti on pathogenicity test of lasiodiplodia theobromae isolates from six host plants and their phylogeny analsysis. The last day was closing session; there were four keynote sessions and all participants were listening together at the same hall. The first keynote speech in first session was Dasuki Amsir, President Director of Perkebunan Nusantara Holding Company on downstream industry development. The second keynote speaker in second session was Elia Massa Manik, President Director PT Pertamina on upstream industry development. The third session was presented by third and fourth speakers on plantation under climate change. The speakers were Ir. Dian Rachmawan, Director of Enterprise & Business Service PT TELKOM, tbk and Prof. Dr. Meine van Noordwijk from ICRAF/ Univ. Wageningen-Netherland. The last session was presented by Dr. Vincent Petiard, Nature Resources Genetics, USA and Maurits Bruggink, Managing Director Ridens Public Affairs on their views of global plantations industry. The closing ceremony was inaugurated by Minister of Agriculture of the Republic Indonesia.